# Patent

## Title
Isoprene synthase variants for improved microbial production of isoprene

## Abstract

The present invention provides methods and compositions comprising at least one isoprene synthase enzyme with improved catalytic activity and/or solubility. In particular, the present invention provides variant plant isoprene synthases for increased isoprene production in microbial host cells. Biosynthetically produced isoprene of the present invention finds use in the manufacture of rubber and elastomers.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation of U.S. patent application Ser. No. 12/429,143 filed Apr. 23, 2009, which is now U.S. Pat. No. 8,173,410, which claims the benefit of U.S. provisional application Ser. No. 61/125,336 filed Apr. 23, 2008, which are hereby incorporated by reference in their entirety.


FIELD OF THE INVENTION
The present invention provides methods and compositions comprising at least one isoprene synthase enzyme with improved catalytic activity and/or solubility. In particular, the present invention provides variant plant isoprene synthases for increased isoprene production in microbial host cells. Biosynthetically produced isoprene of the present invention finds use in the manufacture of rubber and elastomers.
BACKGROUND OF THE INVENTION
Isoprenoids are isoprene polymers that find use in pharmaceuticals, neutraceuticals, flavors, fragrances, and rubber products. Natural isoprenoid supplies, however, are limited due to ecological concerns. For this reason, and to provide isoprenoid compositions having fewer impurities and greater uniformity, isoprenoids such as rubber are often produced synthetically.
Isoprene (2-methyl-1,3-butadiene) is a volatile hydrocarbon that is insoluble in water and soluble in alcohol. Commercially viable quantities of isoprene can be obtained by direct isolation from petroleum C5 cracking fractions or by dehydration of C5 isoalkanes or isoalkenes (Weissermel and Arpe, Industrial Organic Chemistry, 4th ed., Wiley-VCH, pp. 117-122, 2003). The C5 skeleton can also be synthesized from smaller subunits. It would be desirable, however, to have a commercially viable method of producing isoprene that was independent of nonrenewable resources.
Biosynthetic production of isoprene occurs by two distinct metabolic pathways (Julsing et al., Appl Microbiol Biotechnol, 75:1377-1384, 2007). In eukaryotes and archae, isoprene is formed via the mevalonate (MVA) pathway, while some eubacteria and higher plants produce isoprene via the methylerythritol phosphate (MEP) pathway. Isoprene emissions from plants are light and temperature-dependent with increases linked to leaf development. An isoprene-producing enzyme, isoprene synthase, has been identified in Aspen trees (Silver and Fall, Plant Physiol, 97:1588-1591, 1991; and Silver and Fall, J Biol Chem, 270:13010-13016, 1995) and is believed to be responsible for the in vivo production of isoprene from whole leaves. Bacterial production of isoprene has also been described (Kuzma et al., Curr Microbiol, 30:97-103, 1995; and Wilkins, Chemosphere, 32:1427-1434, 1996), and varies in amount with the phase of bacterial growth and the nutrient content of the culture medium (U.S. Pat. No. 5,849,970 to Fall et al.; and Wagner et al., J Bacteriol, 181:4700-4703, 1999, both herein incorporated by reference in their entirety). The levels of isoprene obtainable through bacterial systems of the prior art, however, are insufficient for commercial uses.
Thus what the art needs is an efficient, large scale, bacterial isoprene production process to provide feedstock for the manufacture of isoprenoids.
All patents, patent applications, articles and publications mentioned herein are hereby expressly incorporated herein by reference.
SUMMARY OF THE INVENTION
The present invention provides methods and compositions comprising at least one isoprene synthase enzyme with improved catalytic activity and/or solubility. In particular, the present invention provides variant plant isoprene synthases for increased isoprene production in microbial host cells. Biosynthetically produced isoprene of the present invention finds use in the manufacture of rubber and elastomers.
Specifically, the present invention provides isolated isoprene synthase variants, wherein the variant comprises a substitution at a position corresponding to one or more residues (one, two, three, four, five, six, seven, eight, nine or ten) of a kudzu isoprene synthase comprising the amino acid sequence set forth in SEQ ID NO: 2. In some embodiments, the isoprene synthase variant is a kudzu (Pueraria sp.) isoprene synthase variant or a poplar (Populus sp.) isoprene synthase variant. In some embodiments, the one or more residues are selected from but not limited to the group consisting of L26, E30, F31, Q33, L35, E36, N37, L39, K40, V41, K43, L44, R61, V62, D63, Q65, K87, E94, N95, L99, D100, N105, K137, E138, G143, E144, N182, L184, K185, G187, N189, T190, P225, H226, K247, T257, E258, M259, D266, N334, D353, S357, I358I, E361, N389, I392, I393, K398, E401, C421, Q423, Q424, E425, D426, H430, L432, R433, S434, D437, R443, L462, E463, H476, N478, D479, Q485, D508, P513, A515, Q532, Y533, L537, G538, R539, Y542, A543, and P557. In some embodiments, the one or more residues are selected from but not limited to the group consisting of P24, N25, Y309, D310, L377, F381, E384, Y399, N402, A403, S406, S407, G409, A411, L413, F449, A456, T457, S458, A459, A460, E461, L462, E463, R464, G465, E466, T467, T468, N469, M523, S527, and Y531. In some embodiments, the one or more residues are selected from but not limited to the group consisting of A20, N21, Y22, Q23, R271, W278, F299, V302, and S408. The present invention also provides an isolated isoprene synthase variant having an A20G substitution in a kudzu isoprene synthase having the amino acid sequence set forth in SEQ ID NO: 2. In a subset of these embodiments, the variant comprises at least two substitutions (two, three, four, five, six, seven, eight, nine or ten), wherein one of the substitutions is an A20G substitution in a kudzu isoprene synthase having the amino acid sequence set forth in SEQ ID NO: 2. The present invention also provides an isolated isoprene synthase variant having an S408D substitution in a kudzu isoprene synthase having the amino acid sequence set forth in SEQ ID NO: 2. In a subset of these embodiments, the variant comprises at least two substitutions (two, three, four, five, six, seven, eight, nine or ten), wherein one of the substitutions is an S408D substitution in a kudzu isoprene synthase having the amino acid sequence set forth in SEQ ID NO: 2. In some preferred embodiments, the isoprene synthase variant has at least one improved property as compared to wild-type isoprene synthase. In some particularly preferred embodiments, the at least one improved property is selected from but not limited to the group consisting of specific activity (production of isoprene from dimethylallyl diphosphate), and solubility.
In addition, the present invention further provides a polynucleotide sequence encoding the isoprene synthase variant. Also provided is an expression vector comprising a polynucleotide sequence encoding the isoprene synthase variant in operable combination with a promoter. In further embodiments, the present invention provides a host cell comprising the expression vector. Also provided is a lysate of the host cell, wherein the lysate further comprises lysozyme. In some embodiments, the lysate has a neutral pH (6.5 to 7.5), while in other embodiments the lysate has a basic pH (above 7.5 and below 9.5). The present invention also provides methods of producing isoprene, comprising: (a) providing host cells comprising the expression vector; and (b) culturing the host cells under conditions suitable for producing isoprene. In some embodiments, the methods further comprise (c) recovering the isoprene. In still further embodiments, the methods further comprise (d) polymerizing the isoprene. The present invention further provides methods of detecting isoprene synthase activity, comprising: (a) culturing host cells comprising the expression vector under conditions suitable for producing the isoprene synthase variant; (b) lysing the host cells with a lysis buffer comprising lysozyme to produce a cell lysate; and (c) detecting isoprene synthase activity in the cell lysate by measuring isoprene production from dimethylallyl diphosphate (DMAPP). In some embodiments, the host is selected from but not limited to the group consisting of gram-positive bacterial cells, gram-negative bacterial cells, filamentous fungal cells, and yeast cells. In some preferred embodiments, the host is selected from but not limited to the group consisting of Escherichia sp. (E. coli), Panteoa sp. (P. citrea), Bacillus sp. (B. subtilis), Yarrowia sp. (Y. lipolytica), and Trichoderma (T. reesei). In some embodiments, the host cells are cultured in a medium that includes a carbon source selected from but not limited to the group consisting of glucose, glycerol, glycerine, dihydroxyacetone, yeast extract, biomass, molasses, sucrose, and oil.
Moreover, the present invention provides methods of detecting isoprene in a plurality of samples (high-throughput screening), comprising: (a) providing: i) a plurality of samples each comprising isoprene synthase; ii) a glass plate comprising a plurality of wells; and iii) a seal for the glass plate; (b) placing the plurality of samples in the plurality of wells of the glass plate; (c) sealing the glass plate with the seal to produce a sealed glass plate having a headspace associated with the sample in each of the plurality of wells; (d) incubating the glass plate under conditions in which the isoprene synthase is active; and (e) detecting isoprene in the headspace. In some embodiments, the isoprene is detected by gas chromatography-mass spectrometry (GC-MS). In some embodiments, the plurality of samples comprise host cells comprising an expression vector comprising a polynucleotide sequence encoding an isoprene synthase variant in operable combination with a promoter. In some embodiments, the plurality of samples comprise a lysate of the host cells, lysozyme, and dimethylallyl diphosphate (DMAPP). In some preferred embodiments, the glass plate is a deep-well glass block. In some preferred embodiments, the plurality of wells comprises at least 24 wells (preferably at least 48 wells, more preferably at least 96 wells, still more preferably at least 192 wells, and most preferably at least 384 wells). In particularly preferred embodiments, the plurality of wells each comprise a volume of 2 ml or less (preferably 2 ml to 0.2 ml).
Additionally the present invention provides a host cell comprising a heterologous polynucleotide sequence encoding an isoprene synthase variant in operable combination with a promoter, wherein the isoprene synthase variant comprises a substitution at a position corresponding to one or more residues (one, two, three, four, five, six, seven, eight, nine or ten) of a kudzu isoprene synthase comprising the amino acid sequence set forth in SEQ ID NO: 2. In some embodiments, the one or more residues are selected from but not limited to the group consisting of L26, E30, F31, Q33, L35, E36, N37, L39, K40, V41, K43, L44, R61, V62, D63, Q65, K87, E94, N95, L99, D100, N105, K137, E138, G143, E144, N182, L184, K185, G187, N189, T190, P225, H226, K247, T257, E258, M259, D266, N334, D353, S357, I358I, E361, N389, I392, I393, K398, E401, C421, Q423, Q424, E425, D426, H430, L432, R433, S434, D437, R443, L462, E463, H476, N478, D479, Q485, D508, P513, A515, Q532, Y533, L537, G538, R539, Y542, A543, and P557. In some embodiments, the one or more residues are selected from but not limited to the group consisting of P24, N25, Y309, D310, L377, F381, E384, Y399, N402, A403, S406, S407, G409, A411, L413, F449, A456, T457, 5458, A459, A460, E461, L462, E463, R464, G465, E466, T467, T468, N469, M523, S527, and Y531. In some embodiments, the one or more residues are selected from but not limited to the group consisting of A20, N21, Y22, Q23, R271, W278, F299, V302, and S408. The present invention also provides an isolated isoprene synthase variant having an A20G substitution and/or an S408D substitution in a kudzu isoprene synthase having the amino acid sequence set forth in SEQ ID NO: 2. In some preferred embodiments, the isoprene synthase variant has at least one improved property as compared to wild-type isoprene synthase. In some particularly preferred embodiments, the at least one improved property is selected from but not limited to the group consisting of specific activity (production of isoprene from dimethylallyl diphosphate), and solubility. In some preferred embodiments, the polynucleotide sequence is contained within a plasmid. In other preferred embodiments, the polynucleotide sequence is integrated into a chromosome of the host cell. In some embodiments, the host is selected from but not limited to the group consisting of gram-positive bacterial cells, gram-negative bacterial cells, filamentous fungal cells, and yeast cells. In some preferred embodiments, the host is selected from but not limited to the group consisting of Escherichia sp. (E. coli), Panteoa sp. (P. citrea), Bacillus sp. (B. subtilis), Yarrowia sp. (Y. lipolytica), and Trichoderma (T. reesei). In some embodiments, the host cells are cultured in a medium that includes a carbon source selected from but not limited to the group consisting of glucose, glycerol, glycerine, dihydroxyacetone, yeast extract, biomass, molasses, sucrose, and oil. In some embodiments, the host cell further comprises a heterologous or native nucleic acid encoding an IDI polypeptide and/or a heterologous or native nucleic acid encoding a DXS polypeptide, sometimes in combination with the native DXP pathway (for example, expression of dxs and idi in E. coli in addition to the native DXP pathway). Alternatively the entire DXP pathway (FIG. 15) maybe expressed on a plasmid or integrated on the chromosome as an operon, with a single promoter controlling expression, or promoters of varying strengths (example GI 1.20, GI 1.5, or GI 1.6) controlling one or more of the individual genes. In some embodiments, the host cell further comprises one or more nucleic acids encoding an IDI polypeptide and a DXS polypeptide, while in some preferred embodiments, one vector encodes the isoprene synthase variant, the IDI polypeptide, and the DXS polypeptide. In some embodiments, the host cell further comprises a heterologous nucleic acid encoding an MVA pathway polypeptide (e.g., an MVA pathway polypeptide from Saccharomyces cerevisia or Enterococcus faecalis). In some embodiments, the host cell further comprises one or more nucleic acids encoding an MVA pathway polypeptide and a DXS polypeptide, while in some preferred embodiments, one vector encodes the isoprene synthase variant, the MVA pathway polypeptide, and the DXS polypeptide. In some preferred embodiments, the host cell further comprises one or more nucleic acids encoding a DXS polypeptide, an IDI polypeptide, or one or more of the rest of the DXP pathway polypeptides, and a MVA pathway polypeptide. In some embodiments, the vector further comprises a selectable marker (e.g., antibiotic resistance nucleic acid). Also provided are methods of producing isoprene, comprising: (a) culturing the host cells under suitable culture conditions for production of isoprene; and (b) producing the isoprene. In some embodiments, the methods further comprise (c) recovering the isoprene. In some preferred embodiments, the methods further comprise (d) polymerizing isoprene. The present invention also provides methods of producing isoprene synthase, comprising: (a) providing: (i) a host cell; and (ii) a nucleic acid encoding an isoprene synthase variant in operable combination with a promoter, wherein the isoprene synthase variant comprises a substitution at a position corresponding to one or more residues (one, two, three, four, five, six, seven, eight, nine or ten) of a kudzu isoprene synthase comprising the amino acid sequence set forth in SEQ ID NO: 2; (b) contacting the host cell with the nucleic acid to produce a transformed host cell; and (c) culturing the transformed host cells under suitable culture conditions for production of isoprene synthase.
In another aspect, the invention provides for isolated poplar isoprene synthase variants. In one embodiment, the variant comprises a truncation in the N-terminal portion of isoprene synthase. In another embodiment, the isoprene synthase variant has an increased specific activity compared to a full length isoprene synthase. In another embodiment, the isoprene synthase is P. alba isoprene synthase of SEQ ID NO:120. In another embodiment, wherein the variant is selected from the group consisting of: an MEA variant (SEQ ID NO:122), an MSV variant (SEQ ID NO:124), an MVS variant (SEQ ID NO:126), an MTE variant (SEQ ID NO:128), an MNV variant (SEQ ID NO:130). In another embodiment, the variant is an MEA variant (SEQ ID NO:122). In another embodiment, the variant is selected from the group consisting of: a TRC (−3) variant (SEQ ID NO:136), a TRC (−4) variant (SEQ ID NO:138), a TRC (−5) variant (SEQ ID NO:140), a TRC (−6) variant (SEQ ID NO:142) and a TRC (−7) variant (SEQ ID NO:144). In another embodiment, the variant is a MET variant of P. tremuloides isoprene synthase (SEQ ID NO:146). In another embodiment, the variant is a MET variant of P. trichocharpa isoprene synthase (SEQ ID NO:148).
In another aspect, the invention provides for isolated poplar isoprene synthase variants, wherein the variant comprises a substitution of one or more amino acid residues of a wild type isoprene synthase; and wherein the isoprene synthase variant has increased isoprene synthase activity compared to a wild type isoprene synthase. In one embodiment, the increased isoprene synthase activity is indicated by a host cell comprising the isoprene variant growing at a faster rate in the presence of dimethylallyl pyrophosphate (DMAPP) compared to a host cell comprising a parent isoprene synthase. In another embodiment, the isoprene synthase is the P. alba isoprene synthase of SEQ ID NO:120. In another embodiment, the variant comprises one of more amino acid substitutions selected from the group consisting of V10M, F12S, T15A, E18G, V58I, V58F, L70Q, L70V, L70T, T71P, V79L, E89D, G94A, S119F, F120L, G127R, E175V, T212I, S257A, R262G, A266G, F280L, N297K, F305L, L319M, E323K, A328T, D342E, A359T, K366N, E368D, L374M, S396T, V418S, K438N, H440R, T442I, T442A, I449V, A469S, K500R, K505Q, G507S, S509N, F511Y, and N532K. In another embodiment, at least one amino acid substitution is a L70R substitution. In another embodiment, the variant comprises one of more amino acid substitutions selected from the group consisting of G127R/F511Y, L70Q/G94A/R262G/F305L, F12S/T15A/E18G/N297K, S396T/T442I, V10M/E323K, F120L/A266G, K438N/K500R, V79L/S509N, E175V/S257A/E368D/A469S, T71P/L374M, F280L/H440R, E89D/H440R, V58F/A328T/N532K, S119F/D342E/I449V, and K366N/G507S.
In another aspect, the invention provides for a crystalline form of a polypeptide comprising the amino acid residues of SEQ ID NO:120 (FIG. 19).
In another aspect, the invention provides for methods of producing isoprene, comprising: (a) providing a host cell comprising an expression vector comprising a polynucleotide sequence encoding an isoprene synthase variant; and (b) culturing the host cell under conditions suitable for producing isoprene. In one embodiment, the method further comprises (c) recovering the isoprene. In another embodiment, the method further comprises (d) polymerizing the isoprene.
In another aspect, the invention provides for methods of detecting isoprene synthase activity, comprising: (a) culturing a host cell comprising the expression vector under conditions suitable for producing an isoprene synthase variant; (b) lysing the host cells with a lysis buffer comprising lysozyme to produce a cell lysate; and (c) detecting isoprene synthase activity in the cell lysate by measuring isoprene production from dimethylallyl diphosphate (DMAPP). In one embodiment, the host cell is selected from the group consisting of gram-positive bacterial cells, gram-negative bacterial cells, filamentous fungal cells, and yeast cells. In another embodiment, the host cell is selected from the group consisting of Escherichia sp. (E. coli), Panteoa sp. (P. citrea), Bacillus sp. (B. subtilis), Yarrowia sp. (Y. lipolytica), and Trichoderma (T. reesei). In another embodiment, the host cell is cultured in a medium that includes a carbon source selected from the group consisting of glucose, glycerol, glycerine, dihydroxyacetone, yeast extract, biomass, molasses, sucrose, and oil.
In another aspect, the invention provides for host cells comprising a heterologous polynucleotide sequence encoding an isoprene synthase variant in operable combination with a promoter, wherein the isoprene synthase variant comprises a substitution at a position corresponding to one or more residues (one, two, three, four, five, six, seven, eight, nine or ten) of a poplar isoprene synthase. In one embodiment, the isoprene synthase is the P. alba isoprene synthase of SEQ ID NO:120. In another embodiment, the variant is selected from the group consisting of: an MEA variant (SEQ ID NO:122), an MSV variant (SEQ ID NO:124), an MVS variant (SEQ ID NO:126), an MTE variant (SEQ ID NO:128), an MNV variant (SEQ ID NO:130). In another embodiment, the variant is selected from the group consisting of: a TRC (−3) variant (SEQ ID NO:136), a TRC (−4) variant (SEQ ID NO:138), a TRC (−5) variant (SEQ ID NO:140), a TRC (−6) variant (SEQ ID NO:142) and a TRC (−7) variant (SEQ ID NO:144). In another embodiment, the variant is a MET variant of P. tremuloides isoprene synthase (SEQ ID NO:146). In another embodiment, the variant is a MET variant of P. trichocharpa isoprene synthase (SEQ ID NO:148). In another embodiment, the variant comprises one of more amino acid substitutions selected from the group consisting of V10M, F12S, T15A, E18G, V58I, V58F, L70Q, L70V, L70T, T71P, V79L, E89D, G94A, S119F, F120L, G127R, E175V, T212I, S257A, R262G, A266G, F280L, N297K, F305L, L319M, E323K, A328T, D342E, A359T, K366N, E368D, L374M, S396T, V418S, K438N, H440R, T442I, T442A, I449V, A469S, K500R, K505Q, G507S, S509N, F511Y, and N532K. In another embodiment, at least one amino acid substitution is a L70R substitution. In another embodiment, the variant comprises one of more amino acid substitutions selected from the group consisting of G127R/F511Y, L70Q/G94A/R262G/F305L, F12S/T15A/E18G/N297K, S396T/T442I, V10M/E323K, F120L/A266G, K438N/K500R, V79L/S509N, E175V/S257A/E368D/A469S, T71P/L374M, F280L/H440R, E89D/H440R, V58F/A328T/N532K, S119F/D342E/I449V, and K366N/G507S. In another embodiment, the polynucleotide sequence is contained within a plasmid. In another embodiment, the polynucleotide sequence is integrated into a chromosome of the host cell. In another embodiment, the host is selected from the group consisting of gram-positive bacterial cells, gram-negative bacterial cells, filamentous fungal cells, and yeast cells. In another embodiment, the host is selected from the group consisting of Escherichia sp. (E. coli), Panteoa sp. (P. citrea), Bacillus sp. (B. subtilis), Yarrowia sp. (Y. lipolytica), and Trichoderma (T. reesei). In another embodiment, the host cell is cultured in a medium comprising a carbon source selected from the group consisting of glucose, glycerol, glycerine, dihydroxyacetone, yeast extract, biomass, molasses, sucrose, and oil. In another embodiment, the host cell further comprises a heterologous or native nucleic acid encoding an IDI polypeptide and/or a heterologous or native nucleic acid encoding a DXS polypeptide, optionally in combination with the native DXP pathway. In another embodiment, the host cell further comprises one or more nucleic acids encoding an IDI polypeptide and a DXS polypeptide. In another embodiment, the host cell comprises one vector encoding the isoprene synthase variant, the IDI polypeptide, and the DXS polypeptide. In another embodiment, the host cell further comprises a heterologous nucleic acid encoding an MVA pathway polypeptide selected from the group consisting of an MVA pathway polypeptide from Saccharomyces cerevisia and Enterococcus faecalis. In another embodiment, the host cell further comprises one or more nucleic acids encoding an MVA pathway polypeptide and a DXS polypeptide and wherein one vector encodes the isoprene synthase variant, the MVA pathway polypeptide, and the DXS polypeptide. In another embodiment, the host cell further comprises one or more nucleic acids encoding a DXS polypeptide, an IDI polypeptide, or one or more of the rest of the DXP pathway polypeptides, and a MVA pathway polypeptide.
In another aspect, the invention provides for methods of producing isoprene, comprising: (a) culturing the host cells comprising a heterologous polynucleotide sequence encoding an isoprene synthase variant in operable combination with a promoter, wherein the isoprene synthase variant comprises a substitution at a position corresponding to one or more residues (one, two, three, four, five, six, seven, eight, nine or ten) of a poplar isoprene synthase under suitable culture conditions for production of isoprene; and (b) producing the isoprene. In one embodiment, the method further comprises (c) recovering the isoprene. In another embodiment, the method further comprises (d) polymerizing isoprene.
In another aspect, the invention provides for methods of producing isoprene synthase, comprising: (a) providing: (i) a host cell; and (ii) a nucleic acid encoding an isoprene synthase variant in operable combination with a promoter, wherein the isoprene synthase variant comprises a substitution at a position corresponding to one or more residues (one, two, three, four, five, six, seven, eight, nine or ten) of a P. alba isoprene synthase of SEQ ID NO:120; (b) contacting the host cell with the nucleic acid to produce a transformed host cell; and (c) culturing the transformed host cells under suitable culture conditions for production of isoprene synthase.



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 provides the coding sequence (SEQ ID NO:1) of kudzu (Pueraria montana) isoprene synthase, codon-optimized for expression in Escherichia coli. 
FIG. 2 provides the amino acid sequence (SEQ ID NO:2) of kudzu isoprene synthase.
FIG. 3 provides the coding sequence (SEQ ID NO:6) of poplar (Populus alba x tremula) isoprene synthase, codon-optimized for expression in Escherichia coli. 
FIG. 4 provides the amino acid sequence (SEQ ID NO:7) of poplar (Populus alba x tremula) isoprene synthase.
FIG. 5 provides a kudzu isoprene synthase homology model with the cysteine residues highlighted as space filling molecules.
FIG. 6 provides a poplar isoprene synthase homology model with the cysteine residues highlighted as space filling molecules (dark grey). In addition, the cysteine residues from the kudzu model of FIG. 5 are superimposed on the poplar model as space filling molecules (light grey).
FIG. 7 provides a graph showing the growth curves of the kudzu IspS cysteine mutants of Example 5.
FIG. 8 shows an SDS-PAGE analysis of kudzu IspS cysteine mutants from lysed cells. Pellet and supernatant fractions were prepared by centrifugation.
FIG. 9 provides a map of plasmid MCM93 (pCR2.1-Kudzu).
FIG. 10 provides the nucleotide sequence of plasmid MCM93 (SEQ ID NO:22).
FIG. 11 provides a map of pET24D-Kudzu.
FIG. 12 provides the nucleotide sequence of pET24D-Kudzu (SEQ ID NO:23).
FIG. 13 shows an SDS-PAGE analysis of kudzu isoprene synthase-containing inclusion bodies. Lane M contains molecular weight markers, while the other lanes contain increasing amounts of the purified inclusion body preparation. The kudzu isoprene synthase was estimated to have a purity of >90%.
FIG. 14 provides graphs showing isoprene synthase activity of kudzu site evaluation library (SEL) members for positions Y22, A20 and S408. Most members show highly decreased activity, while conservative substitutions show a lesser decrease in activity. Panel A shows assay results for the Y22 library members in comparison with independent wild type samples (circled WT). Panel B shows assay results for the A20 library members in comparison to wild type samples (circled WT). Panel C shows assay results for the S408 library members, indicating that member S408D has 1.5 to 2-fold higher activity than the average of the wild type controls.
FIG. 15 shows the MVA and DXP metabolic pathways for isoprene (based on F. Bouvier et al., Progress in Lipid Res. 44: 357-429, 2005). The following description includes alternative names for each polypeptide in the pathways and a reference that discloses an assay for measuring the activity of the indicated polypeptide (each of these references are each hereby incorporated by reference in their entireties, particularly with respect to assays for polypeptide activity for polypeptides in the MVA and DXP pathways). Mevalonate Pathway: AACT; Acetyl-CoA acetyltransferase, MvaE, EC 2.3.1.9. Assay: J. Bacteriol., 184: 2116-2122, 2002; HMGS; Hydroxymethylglutaryl-CoA synthase, MvaS, EC 2.3.3.10. Assay: J. Bacteriol., 184: 4065-4070, 2002; HMGR; 3-Hydroxy-3-methylglutaryl-CoA reductase, MvaE, EC 1.1.1.34. Assay: J. Bacteriol., 184: 2116-2122, 2002; MVK; Mevalonate kinase, ERG12, EC 2.7.1.36. Assay: Curr Genet. 19:9-14, 1991. PMK; Phosphomevalonate kinase, ERGS, EC 2.7.4.2, Assay: Mol Cell Biol., 11:620-631, 1991; DPMDC; Diphosphomevalonate decarboxylase, MVD1, EC 4.1.1.33. Assay: Biochemistry, 33:13355-13362, 1994; IDI; Isopentenyl-diphosphate delta-isomerase, IDI1, EC 5.3.3.2. Assay: J. Biol. Chem. 264:19169-19175, 1989. DXP Pathway: DXS; 1-Deoxyxylulose-5-phosphate synthase, dxs, EC 2.2.1.7. Assay: PNAS, 94:12857-62, 1997; DXR; 1-Deoxy-D-xylulose 5-phosphate reductoisomerase, dxr, EC 2.2.1.7. Assay: Eur. J. Biochem. 269:4446-4457, 2002; MCT; 4-Diphosphocytidyl-2C-methyl-D-erythritol synthase, IspD, EC 2.7.7.60. Assay: PNAS, 97: 6451-6456, 2000; CMK; 4-Diphosphocytidyl-2-C-methyl-D-erythritol kinase, IspE, EC 2.7.1.148. Assay: PNAS, 97:1062-1067, 2000; MCS; 2C-Methyl-D-erythritol 2,4-cyclodiphosphate synthase, IspF, EC 4.6.1.12. Assay: PNAS, 96:11758-11763, 1999; HDS; 1-Hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate synthase, ispG, EC 1.17.4.3. Assay: J. Org. Chem., 70:9168-9174, 2005; HDR; 1-Hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase, IspH, EC 1.17.1.2. Assay: JACS, 126:12847-12855, 2004.
FIG. 16 provides a map of pDu27.
FIG. 17 provides the amino acid sequence (SEQ ID NO:118) of 6Xhis N-terminally tagged P. alba IspS in pDu27.
FIG. 18 provides the nucleotide sequence (SEQ ID NO:119) of plasmid pDu27.
FIG. 19 provides the amino acid sequence (SEQ ID NO:120) of full length P. alba IspS in pET24a. Underlined residues indicate the locations of N-terminal truncations in IspS in plasmids pDu39 through pDu43.
FIG. 20 provides the nucleotide sequence (SEQ ID NO:121) of plasmid P. alba pET24a.
FIG. 21 shows purified IspS displays a lower molecular weight “doublet” by SDS-PAGE analysis.
FIG. 22 shows tryptic peptides identified by mass spectrometry.
FIG. 23 provides maps of pDu40, pDu41, pDu42 and pDu43 harboring N-terminal truncations of P. alba IspS.
FIG. 24 provides a map of Pdu39 which is pET24a-P. alba MEA untagged (in strain MD09-173).
FIG. 25 provides the amino acid sequence (SEQ ID NO:122) of truncated “MEA” variant of P. alba IspS in pDu39.
FIG. 26 provides the nucleotide sequence (SEQ ID NO:123) of plasmid pDu39.
FIG. 27 provides the amino acid sequence (SEQ ID NO:124) of truncated “MSV” variant P. alba IspS in pDu41.
FIG. 28 provides the nucleotide sequence (SEQ ID NO:125) of plasmid pDu41 (pET24a-P.alba (MSV) Untagged).
FIG. 29 provides the amino acid sequence (SEQ ID NO:126) of truncated “MVS” variant P. alba IspS in pDu43.
FIG. 30 provides the nucleotide sequence (SEQ ID NO:127) of plasmid pDu43 (pET24a-P.alba (MVS) Untagged).
FIG. 31 provides the amino acid sequence (SEQ ID NO:128) of truncated “MTE” variant of P. alba IspS in pDu42.
FIG. 32 provides the nucleotide sequence (SEQ ID NO:129) of plasmid pDu42 (pET24a-P.alba (MTE) Untagged).
FIG. 33 provides the amino acid sequence (SEQ ID NO:130) of truncated “MNV” P. alba IspS in pDu40.
FIG. 34 provides the nucleotide sequence (SEQ ID NO:131) of plasmid pDu40 (pET24a-P.alba (MNV) Untagged).
FIG. 35 provides maps of MD09-161 and MD09-163, C-terminally TEV, 6×His-tagged IspS variants.
FIG. 36 provides the amino acid sequence (SEQ ID NO:132) of P. alba MEA(+)TEV in MD09-163.
FIG. 37 provides the nucleotide sequence (SEQ ID NO:133) of plasmid MD09-163 (pET24a-P.alba MEA(+)TEV. CDS is underlined, TEV protease site is bold.
FIG. 38 provides the amino acid sequence (SEQ ID NO:134) of P. alba FL(+)TEV in MD09-161.
FIG. 39 provides the nucleotide sequence (SEQ ID NO:135) of plasmid MD09-161 (pET24a-P.alba FL(+)TEV. CDS is underlined, TEV protease site is bold.
FIG. 40 shows a graph representing specific activities of MD09-167, Full length (FL), MD09-165, and Truncated isoprene synthase (MD09-173). Reactions were run at 30° C. for 15 minutes in a solution containing 100 mM Tris, 100 mM NaCl, 50 mM MgCl2, 5 mM DMAPP, and 2.5-4.5 μg isoprene synthase in the supernatant of whole cell lysate.
FIG. 41 shows graphs demonstrating Rate/[E] vs. [DMAPP]. X's represent MD09-173, circles represent MD09-167, diamonds represent MD09-165 and squares represent full length IspS.
FIG. 42 shows a graph showing Isoprene synthase activity vs. [DMAPP]. X's represent data generated with MD09-173 truncated isoprene synthase. Circles represent data generated with MD09-167 isoprene synthase. Diamonds represent data generated with MD09-165 isoprene synthase. Squares represent data generated with full length isoprene synthase. Each data set was run in triplicate from independently grown cultures.
FIG. 43 shows graphs demonstrating the effects of varying kcat and KM and Ki on reaction rate. In panel A, line 1 represents the rate equation of truncated isoprene synthase activity divided by the rate equation of full length isoprene synthase plotted at varying DMAPP concentrations. Line 2 represents the rate equation of full length isoprene synthase in which the kcat has been substituted by the kcat of the truncated isoprene synthase divided by the rate equation of the full length isoprene synthase. Line 3 represents the rate equation of full length isoprene synthase in which the KM has been substituted by the KM of the truncated isoprene synthase divided by the rate equation of the full length isoprene synthase. Line 4 represents the rate equation of full length isoprene synthase in which the Ki has been substituted by the Ki of the truncated isoprene synthase divided by the rate equation of the full length isoprene synthase. Panel B shows a graph demonstrating data fit to the ratio of the rate equation of truncated isoprene synthase to full length isoprene synthase vs. [DMAPP].
FIG. 44 shows a graph demonstrating growth inhibition of MCM531 by Mevalonic Acid (MVA). Cells were grown in TM3 medium in a microtiter plate with different concentrations of MVA. OD600 of quadruplicate wells was measured at the indicated times.
FIG. 45 shows graphs demonstrating DMAPP assays of L70 SSL plate. Dark bars represent either the full length (P. alba pET24a) or pDU39 (truncated) controls. The variants in wells C3 (27), D3 (39), or E3 (51) were chosen for further analysis.
FIG. 46 shows a graph demonstrating the average specific activity of all variants selected for DMAPP assay with protein determination. Error bars show one standard deviation. All 3 L70R variants display higher activity than the control (WT).
FIG. 47 shows a graph demonstrating the average specific activity of all 3 L70R variants compared to the “MEA” truncated P. alba IspS enzyme. Error bars show one standard deviation.
FIG. 48 provides maps of plasmids pDu47-3, pDu47-4, and pDu47-5.
FIG. 49 provides maps of plasmids pDu47-6, pDu47-7, and pDu48.
FIG. 50 provides maps of plasmids pDu49, pDu50 and pDu50-4.
FIG. 51 provides the amino acid sequence (SEQ ID NO:136) of P. alba TRC (−3) in pDu47-3.
FIG. 52 provides the nucleotide sequence (SEQ ID NO:137) of plasmid pDu47-3.
FIG. 53 provides the amino acid sequence (SEQ ID NO:138) of P. alba TRC (−4) in pDu47-4.
FIG. 54 provides the nucleotide sequence (SEQ ID NO:139) of plasmid pDu47-4.
FIG. 55 provides the amino acid sequence (SEQ ID NO:140) of P. alba TRC (−5) in pDu47-5.
FIG. 56 provides the nucleotide sequence (SEQ ID NO:141) of plasmid pDu47-5.
FIG. 57 provides the amino acid sequence (SEQ ID NO:142) of P. alba TRC (−6) in pDu47-6.
FIG. 58 provides the nucleotide sequence (SEQ ID NO:143) of plasmid pDu47-6.
FIG. 59 provides the amino acid sequence (SEQ ID NO:144) of P. alba TRC (−7) in pDu47-7.
FIG. 60 provides the nucleotide sequence (SEQ ID NO:145) of plasmid pDu47-7.
FIG. 61 provides the amino acid sequence (SEQ ID NO:146) of P. tremuloides TRC (MET) in pDu48.
FIG. 62 provides the nucleotide sequence (SEQ ID NO:147) of plasmid pDu48.
FIG. 63 provides the amino acid sequence (SEQ ID NO:148) of P. trichocarpa (TRC) in pDu49.
FIG. 64 provides the nucleotide sequence (SEQ ID NO:149) of plasmid pDu49.
FIG. 65 provides the amino acid sequence (SEQ ID NO:150) of Kudzu TRC (MEA) in pDu50.
FIG. 66 provides the nucleotide sequence (SEQ ID NO:151) of plasmid pDu50.
FIG. 67 provides the amino acid sequence (SEQ ID NO:152) of KudzuTRC (−4) in pDu50-4.
FIG. 68 provides the nucleotide sequence (SEQ ID NO:153) of plasmid pDu50-4.
FIG. 69 shows graphs demonstrating raw and OD-normalized data from DMAPP assay of truncated variants of IspS.
FIG. 70 shows a graph representing the specific activity of IspS truncations. P. alba, P. tremuloides and P. trichocharpa truncations were compared for specific activity relative to the P. alba “full length” variant.
FIG. 71 provides a map of plasmid p9795.
FIG. 72 provides the nucleotide sequence of plasmid p9795 (SEQ ID NO:154).
FIG. 73 provides a map of plasmid pTrcKudzu.
FIG. 74 provides the nucleotide sequence (SEQ ID NO:155) of plasmid pTrcKudzu.
FIG. 75 provides a map of plasmid pMAL-C4X.
FIG. 76 provides the nucleotide sequence (SEQ ID NO:156) of plasmid pMAL-C4X.
FIG. 77 provides a map of plasmid pMAL-C4X-Kudzu.
FIG. 78 provides the nucleotide sequence (SEQ ID NO:157) of plasmid pMAL-C4X-Kudzu.
FIG. 79 provides maps of plasmids pET24 P. tremuloides pET24a and P. trichocharpa pET24a.
FIG. 80 provides the amino acid sequence (SEQ ID NO:158) of P. tremuloides IspS in P. trichocharpa pET24a.
FIG. 81 provides the nucleotide sequence (SEQ ID NO:159) of plasmid P. tremuloides pET24a.
FIG. 82 provides the amino acid sequence (SEQ ID NO:160) of P. trichocharpa IspS in P. trichocharpa pET24a.
FIG. 83 provides the nucleotide sequence (SEQ ID NO:161) of plasmid P. trichocharpa pET24a.
FIG. 84 provides maps of plasmids pDu30, pDu31, and pDu32.
FIG. 85 provides the amino acid sequence (SEQ ID NO:162) of IspS variant P.albaTRC-pET200 in pDu30.
FIG. 86 provides the nucleotide sequence (SEQ ID NO:163) of pDu30.
FIG. 87 provides the amino acid sequence (SEQ ID NO:164) of IspS variant P.tremTRC-pET200 in pDu31.
FIG. 88 provides the nucleotide sequence (SEQ ID NO:165) of pDu31.
FIG. 89 provides the amino acid sequence (SEQ ID NO:166) of IspS variant P.trichTRC-pET200 in pDu32.
FIG. 90 provides the nucleotide sequence (SEQ ID NO:167) of pDu32.
FIG. 91 provides the three-dimensional structure of P. tremuloides IspS shown as a dimer. Chain A is in dark gray, chain B is in medium gray and the single magnesium ion found in each active site is light gray.
FIG. 92 provides a monomer view of the structure of P. tremuloides IspS. The magnesium is shown as a light gray sphere and the N- and C-terminals are indicated.
FIG. 93 shows the structural alignments between (A) BdpS and LS, (B) BdpS and poplar IspS, (C) LS and poplar IspS, and (D) TEAS and poplar IspS. In each case the first structure is in light gray and the second is in dark gray. Divalent cations are shown as spheres.
FIG. 94 shows the three dimensional structure of loops in BdpS and LS. Panel A shows the N-terminal loop of Ls in light gray and the N-terminal loop of BdpS in dark gray. Panel B shows that Loop I and Loop II are structurally homologous.
FIG. 95 shows the N-terminal loop of BdpS (dark gray) and poplar IspS (light gray) are structurally divergent. Panel A shows the N-terminal loop and panel B shows Loop I and Loop II.
FIG. 96 shows the N-terminal loop of LS (light gray) and poplar IspS (dark gray) are structurally divergent. Panel A shows the N-terminal loop and panel B shows Loop I and Loop II.
FIG. 97 shows the N-terminal loop of TEAS (light gray) and poplar IspS (dark gray) are structurally divergent. Panel A shows the N-terminal loop and panel B shows Loop I and Loop II. Loop I is disordered in TEAS.



GENERAL DESCRIPTION OF THE INVENTION
The present invention provides methods and compositions comprising at least one isoprene synthase enzyme with improved catalytic activity and/or solubility. In particular, the present invention provides variant plant isoprene synthases for increased isoprene production in microbial host cells. Biosynthetically produced isoprene of the present invention finds use in the manufacture of rubber and elastomers.
Unless otherwise indicated, the practice of the present invention involves conventional techniques commonly used in molecular biology, microbiology, and recombinant DNA, which are within the skill of the art. Such techniques are known to those of skill in the art and are described in numerous texts and reference works (See e.g., Sambrook et al., “Molecular Cloning: A Laboratory Manual,” Second Edition, Cold Spring Harbor, 1989; and Ausubel et al., “Current Protocols in Molecular Biology,” 1987).
Unless defined otherwise herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. For example, Singleton and Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d Ed., John Wiley and Sons, NY (1994); and Hale and Marham, The Harper Collins Dictionary of Biology, Harper Perennial, NY (1991) provide those of skill in the art with a general dictionaries of many of the terms used in the invention. Although any methods and materials similar or equivalent to those described herein find use in the practice of the present invention, the preferred methods and materials are described herein. Accordingly, the terms defined immediately below are more fully described by reference to the Specification as a whole.
Furthermore, the headings provided herein are not limitations of the various aspects or embodiments of the invention, which can be had by reference to the specification as a whole. Accordingly, the terms defined immediately below are more fully defined by reference to the specification as a whole. Nonetheless, in order to facilitate understanding of the invention, a number of terms are defined below.
Definitions
Unless defined otherwise herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although any methods and materials similar or equivalent to those described herein find use in the practice of the present invention, the preferred methods and materials are described herein. Accordingly, the terms defined immediately below are more fully described by reference to the Specification as a whole.
As used herein, the singular terms “a,” “an,” and “the” include the plural reference unless the context clearly indicates otherwise. Unless otherwise indicated, nucleic acids are written left to right in 5′ to 3′ orientation; amino acid sequences are written left to right in amino to carboxy orientation, respectively. It is to be understood that this invention is not limited to the particular methodology, protocols, and reagents described, as these may vary, depending upon the context they are used by those of skill in the art.
It is intended that every maximum numerical limitation given throughout this specification includes every lower numerical limitation, as if such lower numerical limitations were expressly written herein. Every minimum numerical limitation given throughout this specification will include every higher numerical limitation, as if such higher numerical limitations were expressly written herein. Every numerical range given throughout this specification will include every narrower numerical range that falls within such broader numerical range, as if such narrower numerical ranges were all expressly written herein.
All documents cited are, in relevant part, incorporated herein by reference. However, the citation of any document is not to be construed as an admission that it is prior art with respect to the present invention.
As used herein, the term 2-methyl-1,3-butadiene (CAS#78-79-5) (“isoprene”) refers to the direct and final volatile C5 hydrocarbon product from the elimination of pyrophosphate from 3,3-dimethylallyl pyrophosphate (DMAPP), and does not involve the linking or polymerization of [an] IPP molecule(s) to [a] DMAPP molecule(s). As used herein, the terms “isoprene synthase,” and “IspS,” refer to the enzymes that catalyze the elimination or pyrophosphate from diemethylallyl diphosphate (DMAPP) to form isoprene. In some preferred embodiments, the IspS is an enzyme obtained from plants such as kudzu, poplar or red oak. In some embodiments, the term “IspS” refers to a naturally occurring mature enzyme or portion thereof.
Related (and derivative) proteins comprise “variant proteins.” In some preferred embodiments, variant proteins differ from a parent protein (e.g., kudzu IspS set forth as SEQ ID NO:2 or poplar IspS) and one another by a small number of amino acid residues. The number of differing amino acid residues may be one or more, preferably 1, 2, 3, 4, 5, 10, 15, 20, 30, 40, 50, or more amino acid residues. In some preferred embodiments, the number of different amino acids between variants is between 1 and 10. In some particularly preferred embodiments, related proteins and particularly variant proteins comprise at least 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% amino acid sequence identity. Additionally, a related protein or a variant protein as used herein refers to a protein that differs from another related protein or a parent protein in the number of prominent regions. For example, in some embodiments, variant proteins have 1, 2, 3, 4, 5, or 10 corresponding prominent regions that differ from the parent protein.
Several methods are known in the art that are suitable for generating variants of the enzymes of the present invention, including but not limited to site-saturation mutagenesis, scanning mutagenesis, insertional mutagenesis, random mutagenesis, site-directed mutagenesis, and directed-evolution, as well as various other recombinatorial approaches.
Characterization of wild-type and mutant proteins is accomplished via any means or “test” suitable and is preferably based on the assessment of properties of interest. For example one or more of the following properties are assessed in some embodiments of the present invention: pH stability; temperature stability; oxidative stability; proteolytic stability; solubility; Km and/or specific activity of the conversion of DMAPP to isoprene in vitro; Km and/or specific activity of the conversion of DMAPP to isoprene in vivo in the context of a host organism (e.g., E. coli); and expression of enzyme(s) of the DXP pathway and/or the MVA pathway. Indeed, it is contemplated that enzymes having various degrees of stability, solubility, activity, and/or expression level in one or more of test conditions will find use in the present invention.
As used herein the term “gene” refers to a polynucleotide (e.g., a DNA segment) that encodes a polypeptide and includes regions preceding and following the coding regions as well as intervening sequences (introns) between individual coding segments (exons).
As used herein, “homologous genes” refers to a pair of genes from different, but usually related species, which correspond to each other and which are identical or very similar to each other. The term encompasses genes that are separated by speciation (i.e., the development of new species) (e.g., orthologous genes), as well as genes that have been separated by genetic duplication (e.g., paralogous genes).
As used herein, “ortholog” and “orthologous genes” refer to genes in different species that have evolved from a common ancestral gene (i.e., a homologous gene) by speciation. Typically, orthologs retain the same function during the course of evolution. Identification of orthologs finds use in the reliable prediction of gene function in newly sequenced genomes.
As used herein, “paralog” and “paralogous genes” refer to genes that are related by duplication within a genome. While orthologs retain the same function through the course of evolution, paralogs evolve new functions, even though some functions are often related to the original one.
As used herein, “homology” refers to sequence similarity or identity, with identity being preferred. This homology is determined using standard techniques known in the art (See e.g., Smith and Waterman, Adv Appl Math, 2:482, 1981; Needleman and Wunsch, J Mol Biol, 48:443, 1970; Pearson and Lipman, Proc Natl Acad Sci USA, 85:2444, 1988; programs such as GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, Madison, Wis.; and Devereux et al., Nucl Acid Res, 12:387-395, 1984).
As used herein, an “analogous sequence” is one wherein the function of the gene is essentially the same as the gene based on the kudzu isoprene synthase (IspS) or poplar IspS (IspS). Additionally, analogous genes include at least 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% or 100% sequence identity with the sequence of the kudzu isoprene synthase. In additional embodiments more than one of the above properties applies to the sequence. Analogous sequences are determined by known methods of sequence alignment. A commonly used alignment method is BLAST, although as indicated above and below, there are other methods that also find use in aligning sequences.
One example of a useful algorithm is PILEUP. PILEUP creates a multiple sequence alignment from a group of related sequences using progressive, pair-wise alignments. It can also plot a tree showing the clustering relationships used to create the alignment. PILEUP uses a simplification of the progressive alignment method of Feng and Doolittle (Feng and Doolittle, J Mol Evol, 35:351-360, 1987). The method is similar to that described by Higgins and Sharp (Higgins and Sharp, CABIOS 5:151-153, 1989). Useful PILEUP parameters including a default gap weight of 3.00, a default gap length weight of 0.10, and weighted end gaps.
Another example of a useful algorithm is the BLAST algorithm, described by Altschul et al., (Altschul et al., J Mol Biol, 215:403-410, 1990; and Karlin et al., Proc Natl Acad Sci USA, 90:5873-5787, 1993). A particularly useful BLAST program is the WU-BLAST-2 program (See, Altschul et al., Meth Enzymol, 266:460-480, 1996). WU-BLAST-2 uses several search parameters, most of which are set to the default values. The adjustable parameters are set with the following values: overlap span=1, overlap fraction=0.125, word threshold (T)=11. The HSP S and HSP S2 parameters are dynamic values and are established by the program itself depending upon the composition of the particular sequence and composition of the particular database against which the sequence of interest is being searched. However, the values may be adjusted to increase sensitivity. A % amino acid sequence identity value is determined by the number of matching identical residues divided by the total number of residues of the “longer” sequence in the aligned region. The “longer” sequence is the one having the most actual residues in the aligned region (gaps introduced by WU-Blast-2 to maximize the alignment score are ignored).
Thus, “percent (%) nucleic acid sequence identity” is defined as the percentage of nucleotide residues in a candidate sequence that are identical to the nucleotide residues of the starting sequence (i.e., the sequence of interest). A preferred method utilizes the BLASTN module of WU-BLAST-2 set to the default parameters, with overlap span and overlap fraction set to 1 and 0.125, respectively.
As used herein, the term “hybridization” refers to the process by which a strand of nucleic acid joins with a complementary strand through base pairing, as known in the art.
A nucleic acid sequence is considered to be “selectively hybridizable” to a reference nucleic acid sequence if the two sequences specifically hybridize to one another under moderate to high stringency hybridization and wash conditions. Hybridization conditions are based on the melting temperature (Tm) of the nucleic acid binding complex or probe. For example, “maximum stringency” typically occurs at about Tm-5° C. (5° below the Tm of the probe); “high stringency” at about 5-10° C. below the Tm; “intermediate stringency” at about 10-20° C. below the Tm of the probe; and “low stringency” at about 20-25° C. below the Tm. Functionally, maximum stringency conditions may be used to identify sequences having strict identity or near-strict identity with the hybridization probe; while intermediate or low stringency hybridization can be used to identify or detect polynucleotide sequence homologs.
Moderate and high stringency hybridization conditions are well known in the art. An example of high stringency conditions includes hybridization at about 42° C. in 50% formamide, 5× SSC, 5× Denhardt's solution, 0.5% SDS and 100 μg/ml denatured carrier DNA followed by washing two times in 2× SSC and 0.5% SDS at room temperature and two additional times in 0.1× SSC and 0.5% SDS at 42° C. An example of moderate stringent conditions include an overnight incubation at 37° C. in a solution comprising 20% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate and 20 mg/ml denatured sheared salmon sperm DNA, followed by washing the filters in 1× SSC at about 37-50° C. Those of skill in the art know how to adjust the temperature, ionic strength, etc. as necessary to accommodate factors such as probe length and the like.
As used herein, “recombinant” includes reference to a cell or vector, that has been modified by the introduction of a heterologous nucleic acid sequence or that the cell is derived from a cell so modified. Thus, for example, recombinant cells express genes that are not found in identical form within the native (non-recombinant) form of the cell or express native genes that are otherwise abnormally expressed, under expressed or not expressed at all as a result of deliberate human intervention. “Recombination,” “recombining,” and generating a “recombined” nucleic acid are generally the assembly of two or more nucleic acid fragments wherein the assembly gives rise to a chimeric gene.
In a preferred embodiment, mutant DNA sequences are generated with site saturation mutagenesis in at least one codon. In another preferred embodiment, site saturation mutagenesis is performed for two or more codons. In a further embodiment, mutant DNA sequences have more than 50%, more than 55%, more than 60%, more than 65%, more than 70%, more than 75%, more than 80%, more than 85%, more than 90%, more than 95%, or more than 98% homology with the wild-type sequence. In alternative embodiments, mutant DNA is generated in vivo using any known mutagenic procedure such as, for example, radiation, nitrosoguanidine and the like. The desired DNA sequence is then isolated and used in the methods provided herein.
As used herein, the term “target sequence” refers to a DNA sequence in the host cell that encodes the sequence where it is desired for the incoming sequence to be inserted into the host cell genome. In some embodiments, the target sequence encodes a functional wild-type gene or operon, while in other embodiments the target sequence encodes a functional mutant gene or operon, or a non-functional gene or operon.
As used herein, a “flanking sequence” refers to any sequence that is either upstream or downstream of the sequence being discussed (e.g., for genes A-B-C, gene B is flanked by the A and C gene sequences). In a preferred embodiment, the incoming sequence is flanked by a homology box on each side. In another embodiment, the incoming sequence and the homology boxes comprise a unit that is flanked by stuffer sequence on each side. In some embodiments, a flanking sequence is present on only a single side (either 3′ or 5′), but in preferred embodiments, it is on each side of the sequence being flanked. In some embodiments, a flanking sequence is present on only a single side (either 3′ or 5′), while in preferred embodiments, it is present on each side of the sequence being flanked.
As used herein, the term “stuffer sequence” refers to any extra DNA that flanks homology boxes (typically vector sequences). However, the term encompasses any non-homologous DNA sequence. Not to be limited by any theory, a stuffer sequence provides a noncritical target for a cell to initiate DNA uptake.
As used herein, the terms “amplification” and “gene amplification” refer to a process by which specific DNA sequences are disproportionately replicated such that the amplified gene becomes present in a higher copy number than was initially present in the genome. In some embodiments, selection of cells by growth in the presence of a drug (e.g., an inhibitor of an inhibitable enzyme) results in the amplification of either the endogenous gene encoding the gene product required for growth in the presence of the drug or by amplification of exogenous (i.e., input) sequences encoding this gene product, or both.
“Amplification” is a special case of nucleic acid replication involving template specificity. It is to be contrasted with non-specific template replication (i.e., replication that is template-dependent but not dependent on a specific template). Template specificity is here distinguished from fidelity of replication (i.e., synthesis of the proper polynucleotide sequence) and nucleotide (ribo- or deoxyribo-) specificity. Template specificity is frequently described in terms of “target” specificity. Target sequences are “targets” in the sense that they are sought to be sorted out from other nucleic acid. Amplification techniques have been designed primarily for this sorting out.
As used herein, the term “co-amplification” refers to the introduction into a single cell of an amplifiable marker in conjunction with other gene sequences (i.e., comprising one or more non-selectable genes such as those contained within an expression vector) and the application of appropriate selective pressure such that the cell amplifies both the amplifiable marker and the other, non-selectable gene sequences. The amplifiable marker may be physically linked to the other gene sequences or alternatively two separate pieces of DNA, one containing the amplifiable marker and the other containing the non-selectable marker, may be introduced into the same cell.
As used herein, the terms “amplifiable marker,” “amplifiable gene,” and “amplification vector” refer to a gene or a vector encoding a gene, which permits the amplification of that gene under appropriate growth conditions.
“Template specificity” is achieved in most amplification techniques by the choice of enzyme. Amplification enzymes are enzymes that, under conditions they are used, will process only specific sequences of nucleic acid in a heterogeneous mixture of nucleic acid. For example, in the case of Qβ replicase, MDV-1 RNA is the specific template for the replicase (See e.g., Kacian et al., Proc Natl Acad Sci USA 69:3038, 1972) and other nucleic acids are not replicated by this amplification enzyme. Similarly, in the case of T7 RNA polymerase, this amplification enzyme has a stringent specificity for its own promoters (See, Chamberlin et al., Nature 228:227, 1970). In the case of T4 DNA ligase, the enzyme will not ligate the two oligonucleotides or polynucleotides, where there is a mismatch between the oligonucleotide or polynucleotide substrate and the template at the ligation junction (See, Wu and Wallace, Genomics 4:560, 1989). Finally, Taq and Pfu polymerases, by virtue of their ability to function at high temperature, are found to display high specificity for the sequences bounded and thus defined by the primers; the high temperature results in thermodynamic conditions that favor primer hybridization with the target sequences and not hybridization with non-target sequences.
As used herein, the term “amplifiable nucleic acid” refers to nucleic acids, which may be amplified by any amplification method. It is contemplated that “amplifiable nucleic acid” will usually comprise “sample template.”
As used herein, the term “sample template” refers to nucleic acid originating from a sample, which is analyzed for the presence of “target” (defined below). In contrast, “background template” is used in reference to nucleic acid other than sample template, which may or may not be present in a sample. Background template is most often inadvertent. It may be the result of carryover, or it may be due to the presence of nucleic acid contaminants sought to be purified away from the sample. For example, nucleic acids from organisms other than those to be detected may be present as background in a test sample.
As used herein, the term “primer” refers to an oligonucleotide, whether occurring naturally as in a purified restriction digest or produced synthetically, which is capable of acting as a point of initiation of synthesis when placed under conditions in which synthesis of a primer extension product which is complementary to a nucleic acid strand is induced, (i.e., in the presence of nucleotides and an inducing agent such as DNA polymerase and at a suitable temperature and pH). The primer is preferably single stranded for maximum efficiency in amplification, but may alternatively be double stranded. If double stranded, the primer is first treated to separate its strands before being used to prepare extension products. Preferably, the primer is an oligodeoxyribonucleotide. The primer must be sufficiently long to prime the synthesis of extension products in the presence of the inducing agent. The exact lengths of the primers will depend on many factors, including temperature, source of primer and the use of the method.
As used herein, the term “probe” refers to an oligonucleotide (i.e., a sequence of nucleotides), whether occurring naturally as in a purified restriction digest or produced synthetically, recombinantly or by PCR amplification, which is capable of hybridizing to another oligonucleotide of interest. A probe may be single-stranded or double-stranded. Probes are useful in the detection, identification and isolation of particular gene sequences. It is contemplated that any probe used in the present invention will be labeled with any “reporter molecule,” so that is detectable in any detection system, including, but not limited to enzyme (e.g., ELISA, as well as enzyme-based histochemical assays), fluorescent, radioactive, and luminescent systems. It is not intended that the present invention be limited to any particular detection system or label.
As used herein, the term “target,” when used in reference to the polymerase chain reaction, refers to the region of nucleic acid bounded by the primers used for polymerase chain reaction. Thus, the “target” is sought to be sorted out from other nucleic acid sequences. A “segment” is defined as a region of nucleic acid within the target sequence.
As used herein, in one embodiment, the term “polymerase chain reaction” (“PCR”) refers to the methods of U.S. Pat. Nos. 4,683,195 4,683,202, and 4,965,188, hereby incorporated by reference, which include methods for increasing the concentration of a segment of a target sequence in a mixture of genomic DNA without cloning or purification. This process for amplifying the target sequence consists of introducing a large excess of two oligonucleotide primers to the DNA mixture containing the desired target sequence, followed by a precise sequence of thermal cycling in the presence of a DNA polymerase. The two primers are complementary to their respective strands of the double stranded target sequence. To effect amplification, the mixture is denatured and the primers then annealed to their complementary sequences within the target molecule. Following annealing, the primers are extended with a polymerase so as to form a new pair of complementary strands. The steps of denaturation, primer annealing and polymerase extension can be repeated many times (i.e., denaturation, annealing and extension constitute one “cycle”; there can be numerous “cycles”) to obtain a high concentration of an amplified segment of the desired target sequence. The length of the amplified segment of the desired target sequence is determined by the relative positions of the primers with respect to each other, and therefore, this length is a controllable parameter. By virtue of the repeating aspect of the process, the method is referred to as the “polymerase chain reaction” (hereinafter “PCR”). Because the desired amplified segments of the target sequence become the predominant sequences (in terms of concentration) in the mixture, they are said to be “PCR amplified”.
As used herein, the term “amplification reagents” refers to those reagents (deoxyribonucleotide triphosphates, buffer, etc.), needed for amplification except for primers, nucleic acid template and the amplification enzyme. Typically, amplification reagents along with other reaction components are placed and contained in a reaction vessel (test tube, microwell, etc.).
With PCR, it is possible to amplify a single copy of a specific target sequence in genomic DNA to a level detectable by several different methodologies (e.g., hybridization with a labeled probe; incorporation of biotinylated primers followed by avidin-enzyme conjugate detection; incorporation of 32P-labeled deoxynucleotide triphosphates, such as dCTP or dATP, into the amplified segment). In addition to genomic DNA, any oligonucleotide or polynucleotide sequence can be amplified with the appropriate set of primer molecules. In particular, the amplified segments created by the PCR process itself are, themselves, efficient templates for subsequent PCR amplifications.
As used herein, the terms “PCR product,” “PCR fragment,” and “amplification product” refer to the resultant mixture of compounds after two or more cycles of the PCR steps of denaturation, annealing and extension are complete. These terms encompass the case where there has been amplification of one or more segments of one or more target sequences.
As used herein, the term “RT-PCR” refers to the replication and amplification of RNA sequences. In this method, reverse transcription is coupled to PCR, most often using a one enzyme procedure in which a thermostable polymerase is employed, as described in U.S. Pat. No. 5,322,770, herein incorporated by reference. In RT-PCR, the RNA template is converted to cDNA due to the reverse transcriptase activity of the polymerase, and then amplified using the polymerizing activity of the polymerase (i.e., as in other PCR methods).
As used herein, the terms “restriction endonucleases” and “restriction enzymes” refer to bacterial enzymes, each of which cut double-stranded DNA at or near a specific nucleotide sequence.
A “restriction site” refers to a nucleotide sequence recognized and cleaved by a given restriction endonuclease and is frequently the site for insertion of DNA fragments. In certain embodiments of the invention restriction sites are engineered into the selective marker and into 5′ and 3′ ends of the DNA construct.
As used herein, the term “chromosomal integration” refers to the process whereby an incoming sequence is introduced into the chromosome of a host cell. The homologous regions of the transforming DNA align with homologous regions of the chromosome. Subsequently, the sequence between the homology boxes is replaced by the incoming sequence in a double crossover (i.e., homologous recombination). In some embodiments of the present invention, homologous sections of an inactivating chromosomal segment of a DNA construct align with the flanking homologous regions of the indigenous chromosomal region of the Escherichia chromosome. Subsequently, the indigenous chromosomal region is deleted by the DNA construct in a double crossover (i.e., homologous recombination).
“Homologous recombination” means the exchange of DNA fragments between two DNA molecules or paired chromosomes at the site of identical or nearly identical nucleotide sequences. In a preferred embodiment, chromosomal integration is homologous recombination.
“Homologous sequences” as used herein means a nucleic acid or polypeptide sequence having 100%, 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 88%, 85%, 80%, 75%, or 70% sequence identity to another nucleic acid or polypeptide sequence when optimally aligned for comparison. In some embodiments, homologous sequences have between 85% and 100% sequence identity, while in other embodiments there is between 90% and 100% sequence identity, and in more preferred embodiments, there is 95% and 100% sequence identity.
As used herein “amino acid” refers to peptide or protein sequences or portions thereof. The terms “protein,” “peptide,” and “polypeptide” are used interchangeably.
As used herein, the term “heterologous protein” refers to a protein or polypeptide that does not naturally occur in the host cell. Examples of heterologous proteins include enzymes such as isoprene synthases. In some embodiments, the genes encoding the proteins are naturally occurring genes, while in other embodiments mutated and/or synthetic genes are used.
As used herein, “homologous protein” refers to a protein or polypeptide native or naturally occurring in a cell. In preferred embodiments, the cell is a Gram-negative cell, while in particularly preferred embodiments the cell is an Escherichia host cell.
An enzyme is “overexpressed” in a host cell if the enzyme is expressed in the cell at a higher level that the level at which it is expressed in a corresponding wild-type cell.
The terms “protein” and “polypeptide” are used interchangeability herein. The 3-letter code for amino acids as defined in conformity with the IUPAC-IUB Joint Commission on Biochemical Nomenclature (JCBN) is used through out this disclosure. It is also understood that a polypeptide may be coded for by more than one nucleotide sequence due to the degeneracy of the genetic code.
The term “mature” form of a protein or peptide refers to the final functional form of the protein or peptide. To exemplify, a mature form of kudzu isoprene synthase includes the amino acid sequence of SEQ ID NO:2.
The term “precursor” form of a protein or peptide refers to a mature form of the protein having a prosequence operably linked to the amino or carbonyl terminus of the protein. The precursor may also have a “signal” sequence operably linked, to the amino terminus of the prosequence. The precursor may also have additional polynucleotides that are involved in post-translational activity (e.g., polynucleotides cleaved therefrom to leave the mature form of a protein or peptide).
“Naturally occurring enzyme” refers to an enzyme having the unmodified amino acid sequence identical to that found in nature. Naturally occurring enzymes include native enzymes, those enzymes naturally expressed or found in the particular microorganism.
The term “identical” in the context of two nucleic acids or polypeptide sequences refers to the residues in the two sequences that are the same when aligned for maximum correspondence, as measured using one of the following sequence comparison or analysis algorithms.
The term “optimal alignment” refers to the alignment giving the highest percent identity score.
“Percent sequence identity,” “percent amino acid sequence identity,” “percent gene sequence identity,” and/or “percent nucleic acid/polynucloetide sequence identity,” with respect to two amino acid, polynucleotide and/or gene sequences (as appropriate), refer to the percentage of residues that are identical in the two sequences when the sequences are optimally aligned. Thus, 80% amino acid sequence identity means that 80% of the amino acids in two optimally aligned polypeptide sequences are identical.
The phrase “substantially identical” in the context of two nucleic acids or polypeptides thus refers to a polynucleotide or polypeptide that comprising at least 70% sequence identity, preferably at least 75%, preferably at least 80%, preferably at least 85%, preferably at least 90%, preferably at least 95%, preferably at least 97%, preferably at least 98% and preferably at least 99% sequence identity as compared to a reference sequence using the programs or algorithms (e.g., BLAST, ALIGN, CLUSTAL) using standard parameters. One indication that two polypeptides are substantially identical is that the first polypeptide is immunologically cross-reactive with the second polypeptide. Typically, polypeptides that differ by conservative amino acid substitutions are immunologically cross-reactive. Thus, a polypeptide is substantially identical to a second polypeptide, for example, where the two peptides differ only by a conservative substitution. Another indication that two nucleic acid sequences are substantially identical is that the two molecules hybridize to each other under stringent conditions (e.g., within a range of medium to high stringency).
The term “isolated” or “purified” refers to a material that is removed from its original environment (e.g., the natural environment if it is naturally occurring). For example, the material is said to be “purified” when it is present in a particular composition in a higher or lower concentration than exists in a naturally occurring or wild type organism (e.g., kudzu) or in combination with components not normally present upon expression from a naturally occurring or wild type organism. For example, a naturally-occurring polynucleotide or polypeptide present in a living animal is not isolated, but the same polynucleotide or polypeptide, separated from some or all of the coexisting materials in the natural system, is isolated. Such polynucleotides could be part of a vector, and/or such polynucleotides or polypeptides could be part of a composition, and still be isolated in that such vector or composition is not part of its natural environment. In preferred embodiments, a nucleic acid or protein is said to be purified, for example, if it gives rise to essentially one band in an electrophoretic gel or blot.
The term “isolated”, when used in reference to a DNA sequence, refers to a DNA sequence that has been removed from its natural genetic milieu and is thus free of other extraneous or unwanted coding sequences, and is in a form suitable for use within genetically engineered protein production systems. Similarly, the term “isolated”, when used in reference to a recombinant DNA sequence, refers to a DNA sequence that has been removed from the genetic milieu of the host organism and is thus free of other extraneous or unwanted coding sequences (e.g., kudzu IspS expression vector propagated in E. coli). Such isolated molecules are those that are separated from their natural environment and include cDNA and genomic clones. Isolated DNA molecules of the present invention are free of other genes with which they are ordinarily associated, but may include naturally occurring 5′ and 3′ untranslated regions such as promoters and terminators. The identification of associated regions will be evident to one of ordinary skill in the art (See e.g., Dynan and Tijan, Nature 316:774-78, 1985). The term “an isolated DNA sequence” is alternatively referred to as “a cloned DNA sequence”.
The term “isolated,” when used in reference to a protein, refers to a protein that is found in a condition other than its native environment. In a preferred form, the isolated protein is substantially free of other proteins, particularly other homologous proteins. Similarly, the term “isolated”, when used in reference to a recombinantly produced protein, refers to a protein that has been removed from the proteinaceous milieu of the host organism and is thus free of other extraneous or unwanted proteins (e.g., recombinant kudzu IspS produced in E. coli). An isolated protein is more than 10% pure, preferably more than 20% pure, and even more preferably more than 30% pure, as determined by SDS-PAGE. Further aspects of the invention encompass the protein in a highly purified form (i.e., more than 40% pure, more than 60% pure, more than 80% pure, more than 90% pure, more than 95% pure, more than 97% pure, and even more than 99% pure), as determined by SDS-PAGE.
The following cassette mutagenesis method may be used to facilitate the construction of the enzyme variants of the present invention, although other methods may be used. First, as described herein, a naturally-occurring gene encoding the enzyme is obtained and sequenced in whole or in part. Then, the sequence is scanned for a point at which it is desired to make a mutation (deletion, insertion or substitution) of one or more amino acids in the encoded enzyme. The sequences flanking this point are evaluated for the presence of restriction sites for replacing a short segment of the gene with an oligonucleotide pool which when expressed will encode various mutants. Such restriction sites are preferably unique sites within the protein gene so as to facilitate the replacement of the gene segment. However, any convenient restriction site that is not overly redundant in the enzyme gene may be used, provided the gene fragments generated by restriction digestion can be reassembled in proper sequence. If restriction sites are not present at locations within a convenient distance from the selected point (from 10 to 15 nucleotides), such sites are generated by substituting nucleotides in the gene in such a fashion that neither the reading frame nor the amino acids encoded are changed in the final construction. Mutation of the gene in order to change its sequence to conform to the desired sequence is accomplished by M13 primer extension in accord with generally known methods. The task of locating suitable flanking regions and evaluating the needed changes to arrive at two convenient restriction site sequences is made routine by the redundancy of the genetic code, a restriction enzyme map of the gene and the large number of different restriction enzymes. Note that if a convenient flanking restriction site is available, the above method need be used only in connection with the flanking region that does not contain a site.
Once the naturally-occurring DNA and/or synthetic DNA is cloned, the restriction sites flanking the positions to be mutated are digested with the cognate restriction enzymes and a plurality of end termini-complementary oligonucleotide cassettes are ligated into the gene. The mutagenesis is simplified by this method because all of the oligonucleotides can be synthesized so as to have the same restriction sites, and no synthetic linkers are necessary to create the restriction sites.
As used herein, “corresponding to,” refers to a residue at the enumerated position in a protein or peptide, or a residue that is analogous, homologous, or equivalent to an enumerated residue in a protein or peptide. As used herein, “corresponding region,” generally refers to an analogous position along related proteins or a parent protein.
As used herein, the term, “combinatorial mutagenesis” refers to methods in which libraries of variants of a starting sequence are generated. In these libraries, the variants contain one or several mutations chosen from a predefined set of mutations. In addition, the methods provide means to introduce random mutations, which were not members of the predefined set of mutations. In some embodiments, the methods include those set forth in U.S. application Ser. No. 09/699,250, hereby incorporated by reference. In alternative embodiments, combinatorial mutagenesis methods encompass commercially available kits (e.g., QUIKCHANGE Multisite mutagenesis kit, Stratagene, San Diego, Calif.).
As used herein, the term “library of mutants” refers to a population of cells which are identical in most of their genome but include different homologues of one or more genes. Such libraries can be used, for example, to identify genes or operons with improved traits.
As used herein, the terms “starting gene” and “parent gene” refer to a gene of interest that encodes a protein of interest that is to be improved and/or changed using the present invention.
As used herein, the terms “multiple sequence alignment” and “MSA” refer to the sequences of multiple homologs of a starting gene that are aligned using an algorithm (e.g., Clustal W).
As used herein, the terms “consensus sequence” and “canonical sequence” refer to an archetypical amino acid sequence against which all variants of a particular protein or sequence of interest are compared. The terms also refer to a sequence that sets forth the nucleotides that are most often present in a DNA sequence of interest. For each position of a gene, the consensus sequence gives the amino acid that is most abundant in that position in the MSA.
As used herein, the term “consensus mutation” refers to a difference in the sequence of a starting gene and a consensus sequence. Consensus mutations are identified by comparing the sequences of the starting gene and the consensus sequence obtained from a MSA. In some embodiments, consensus mutations are introduced into the starting gene such that it becomes more similar to the consensus sequence. Consensus mutations also include amino acid changes that change an amino acid in a starting gene to an amino acid that is more frequently found in an MSA at that position relative to the frequency of that amino acid in the starting gene. Thus, the term consensus mutation comprises all single amino acid changes that replace an amino acid of the starting gene with an amino acid that is more abundant than the amino acid in the MSA.
The terms “modified sequence” and “modified genes” are used interchangeably herein to refer to a sequence that includes a deletion, insertion or interruption of naturally occurring nucleic acid sequence. In some preferred embodiments, the expression product of the modified sequence is a truncated protein (e.g., if the modification is a deletion or interruption of the sequence). In some particularly preferred embodiments, the truncated protein retains biological activity. In alternative embodiments, the expression product of the modified sequence is an elongated protein (e.g., modifications comprising an insertion into the nucleic acid sequence). In some embodiments, an insertion leads to a truncated protein (e.g., when the insertion results in the formation of a stop codon). Thus, an insertion may result in either a truncated protein or an elongated protein as an expression product.
As used herein, the terms “mutant sequence” and “mutant gene” are used interchangeably and refer to a sequence that has an alteration in at least one codon occurring in a host cell's wild-type sequence. The expression product of the mutant sequence is a protein with an altered amino acid sequence relative to the wild-type. The expression product may have an altered functional capacity (e.g., enhanced enzymatic activity).
The terms “mutagenic primer” or “mutagenic oligonucleotide” (used interchangeably herein) are intended to refer to oligonucleotide compositions which correspond to a portion of the template sequence and which are capable of hybridizing thereto. With respect to mutagenic primers, the primer will not precisely match the template nucleic acid, the mismatch or mismatches in the primer being used to introduce the desired mutation into the nucleic acid library. As used herein, “non-mutagenic primer” or “non-mutagenic oligonucleotide” refers to oligonucleotide compositions that match precisely to the template nucleic acid. In one embodiment of the invention, only mutagenic primers are used. In another preferred embodiment of the invention, the primers are designed so that for at least one region at which a mutagenic primer has been included, there is also non-mutagenic primer included in the oligonucleotide mixture. By adding a mixture of mutagenic primers and non-mutagenic primers corresponding to at least one of the mutagenic primers, it is possible to produce a resulting nucleic acid library in which a variety of combinatorial mutational patterns are presented. For example, if it is desired that some of the members of the mutant nucleic acid library retain their parent sequence at certain positions while other members are mutant at such sites, the non-mutagenic primers provide the ability to obtain a specific level of non-mutant members within the nucleic acid library for a given residue. The methods of the invention employ mutagenic and non-mutagenic oligonucleotides which are generally between 10-50 bases in length, more preferably about 15-45 bases in length. However, it may be necessary to use primers that are either shorter than 10 bases or longer than 50 bases to obtain the mutagenesis result desired. With respect to corresponding mutagenic and non-mutagenic primers, it is not necessary that the corresponding oligonucleotides be of identical length, but only that there is overlap in the region corresponding to the mutation to be added.
Primers may be added in a pre-defined ratio according to the present invention. For example, if it is desired that the resulting library have a significant level of a certain specific mutation and a lesser amount of a different mutation at the same or different site, by adjusting the amount of primer added, it is possible to produce the desired biased library. Alternatively, by adding lesser or greater amounts of non-mutagenic primers, it is possible to adjust the frequency with which the corresponding mutation(s) are produced in the mutant nucleic acid library.
The terms “wild-type sequence” or “wild-type gene” are used interchangeably herein, to refer to a sequence that is native or naturally occurring in a host cell. In some embodiments, the wild-type sequence refers to a sequence of interest that is the starting point of a protein-engineering project. The wild-type sequence may encode either a homologous or heterologous protein. A homologous protein is one the host cell would produce without intervention. A heterologous protein is one that the host cell would not produce but for the intervention.
As used herein the term “lysate” refers to a solution containing the contents of lysed cells. In some embodiments, the lysate is a bacterial cell lysate (e.g., E. coli cells lysed using READYLYSETM lysozyme solution from Epicentre; or E. coli cells lysed using a French Pressure cell).
As used herein the term “lysozyme” refers to a glycosidase that hydrolyzes the bond between N-acetyl muramic acid and N-acetul glucosamine, thus cleaving an important polymer in the cell wall of many bacteria. Suitable lysozymes for use with the present invention include but are not limited to hen egg white lysozyme (Sigma), T4 lysozyme, recombinant non-mammalian, non-avian lysozyme (READYLYSETM), or a fungal lysozyme.
As used herein, the term “headspace” refers to the vapor/air mixture trapped above a solid or liquid sample in a sealed vessel.
As used herein, the terms “high throughput screening” and “HTS” refer to measuring isoprene in at least 96 samples in 4 hours or less. In preferred embodiments, the sample volume is less than 2 mL.
Unless otherwise noted, all component or composition levels are in reference to the active level of that component or composition, and are exclusive of impurities, for example, residual solvents or by-products, which may be present in commercially available sources.
Enzyme components weights are based on total active protein. All percentages and ratios are calculated by weight unless otherwise indicated. All percentages and ratios are calculated based on the total composition unless otherwise.
DETAILED DESCRIPTION OF THE INVENTION
Isoprene monomer is employed in the manufacture of polyisoprene and various copolymers (with isobutylene, butadiene, styrene, or other monomers). To build a strain (prokaryotic or eukaryotic) capable of producing commercially viable levels of isoprene requires optimization of the entire pathway, either MVA to isoprene or DXP to isoprene. A key enzyme in the pathway is isoprene synthase (IspS), which converts the precursor DMAPP to isoprene. The only isoprene synthases (IspS) identified to date are those from plants such as poplar, English oak and kudzu vine. Although some bacteria, such as Bacillus subtilis, also produce isoprene, a prokaryotic IspS has yet to be identified and the native IspS activity in Bacillus is not sufficient for a commercial process. The plant IspS enzymes identified to date have been partially characterized in part by expression in E. coli and some of the kinetic parameters of these enzymes have been determined in vitro with purified protein. However, the kinetic parameters (Km, rate etc) of the native IspS enzymes are insufficient for commercial production of isoprene in a biological host.
To solve this problem as described herein, a plant IspS is expressed in a bacterial host. In addition the IspS is engineered for a change in a property of interest. Characterization of wild-type and mutant IspS is accomplished via any means or “test” suitable and is preferably based on the assessment of properties of interest. Properties of interest include but are not limited to: pH optima, temperature stability (e.g., Tm value), intracellular and extracellular solubility, Km value, kcat value, or specific activity, as well as sensitivity to potential inhibitors including substrate or product inhibition. Oxidative and proteolytic stability are also of interest. Furthermore, activation or inhibition due to metal ion effects and ionic strength is of interest. These properties and parameters can be assessed by the conversion of DMAPP to isoprene in vitro with purified or partially purified isoprene synthase or in vivo in the context of a host organism such as E. coli expressing the DXP pathway, the MVA pathway, or both. It is contemplated that enzymes having various degrees of stability, solubility, activity, and/or expression level in one or more of test conditions will find use in the present invention for the production of isoprene in a diversity of hosts. High throughput methods such as those described in Example 10 are required to investigate these properties in an economical manner.
The invention features compositions and methods for the production of increased amounts of isoprene. In particular, these compositions and methods increase the rate of isoprene production and increase the total amount of isoprene that is produced. The biosynthetic processes for isoprene production described herein are a desirable alternative to using natural rubber. As discussed further below, the amount of isoprene produced by cells can be greatly increased by introducing a heterologous nucleic acid encoding an isoprene synthase (IspS) polypeptide into the cells. Isoprene synthase polypeptides convert dimethylallyl diphosphate (DMAPP) into isoprene. As shown in the examples, a heterologous Pueraria montana (kudzu) isoprene synthase polypeptide and variants thereof was expressed in Gram-negative bacterial cells (e.g., Escherichia coli). Also shown in the examples and contemplated within the scope of the invention are poplar isoprene synthase polypeptide and variants thereof was expressed in Gram-negative bacterial cells (e.g., Escherichia coli).
Heterologous expression of a plant IspS in bacterial host cells resulted in the production of more isoprene than the corresponding cells lacking the plant IspS.
It has been shown that mutating amino-acid residues on the surface of protease enzymes can improve their activity, expression, and stability (WO2008/153925, WO2008/153934, WO2008/153935). Surprisingly, we have found that mutating amino-acid residues on the surface of a completely different enzyme, isoprene synthase, can enhance its expression, solubility, and activity. L70R is an example of such a beneficial surface mutation.
Elucidation of the three-dimensional structure of an enzyme is essential for accurately identifying amino-acid residues on its surface. Homology modeling using structures with sequences approximately 40% identical to isoprene synthase (e.g., bornyl synthase and limonene synthase, the enzymes of known structure with closest identity to isoprene synthase) can reveal gross aspects of the modeled enzyme structure, but is insufficient to precisely identify surface-exposed residues and quantify their degree of surface exposure. Surface exposure of an amino-acid residue is quantified by the percentage of solvent-accessible surface area of its side chain.
The following classes of mutations in isoprene synthase may improve solubility of the enzyme by targeting amino-acid residues that are >50% solvent-exposed, preferably >65% solvent-exposed, and most preferably >85% solvent-exposed:
Hydrophobic→positively charged, and vice versa
Hydrophobic→negatively charged, and vice versa
Hydrophobic→neutral polar, and vice versa
Neutral polar→positively charged, and vice versa
Neutral polar→negatively charged, and vice versa
Positively charged→negatively charged, and vice versa
Additionally isoprene production by cells containing a heterologous isoprene synthase nucleic acid can be enhanced by increasing the amount of a 1-deoxy-D-xylulose-5-phosphate synthase (DXS) polypeptide, and/or an isopentenyl diphosphate isomerase (IDI) polypeptide, expressed by the cells. For example, a DXS nucleic acid and/or an IDI nucleic acid can be introduced into the cells. The DXS nucleic acid may be a heterologous nucleic acid or a duplicate copy of an endogenous nucleic acid. Similarly, the IDI nucleic acid may be a heterologous nucleic acid or a duplicate copy of an endogenous nucleic acid. In some embodiments, the amount of DXS and/or IDI polypeptide is increased by replacing the endogenous DXS and/or IDI promoters or regulatory regions with other promoters and/or regulatory regions that result in greater transcription of the DXS and/or IDI nucleic acids. In some embodiments, the cells contain both a heterologous nucleic acid encoding an isoprene synthase polypeptide (e.g., a plant isoprene synthase nucleic acid) and a duplicate copy of an endogenous nucleic acid encoding an isoprene synthase polypeptide.
The encoded DXS and IDI polypeptides are part of the DXP pathway for the biosynthesis of isoprene (FIG. 15). DXS polypeptides convert pyruvate and D-glyceraldehyde-3-phosphate into 1-deoxy-D-xylulose-5-phosphate. While not intending to be bound by any particular theory, it is believed that increasing the amount of DXS polypeptide increases the flow of carbon through the DXP pathway, leading to greater isoprene production. IDI polypeptides catalyze the interconversion of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). While not intending to be bound by any particular theory, it is believed that increasing the amount of IDI polypeptide in cells increases the amount of IPP that is converted into DMAPP, which in turn is converted into isoprene.
In some embodiments the production of isoprene by cells containing a heterologous isoprene synthase nucleic acid can be augmented by increasing expression of a MVA polypeptide in the cells (FIG. 15). Exemplary MVA pathways polypeptides include any of the following polypeptides: acetyl-CoA acetyltransferase (AA-CoA thiolase) polypeptides, 3-hydroxy-3-methylglutaryl-CoA synthase (HMG-CoA synthase) polypeptides, 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase) polypeptides, mevalonate kinase (MVK) polypeptides, phosphomevalonate kinase (PMK) polypeptides, diphosphomevalonte decarboxylase (MVD) polypeptides, IDI polypeptides, and polypeptides (e.g., fusion polypeptides) having an activity of two or more MVA pathway polypeptides. For example, one or more MVA pathway nucleic acids can be introduced into the cells. In some embodiments, the cells contain the upper MVA pathway, which includes AA-CoA thiolase, HMG-CoA synthase, and HMG-CoA reductase nucleic acids. In some embodiments, the cells contain the lower MVA pathway, which includes MVK, PMK, MVD, and IDI nucleic acids. In some embodiments, the cells contain the entire MVA pathway, which includes AA-CoA thiolase, HMG-CoA synthase, HMG-CoA reductase, MVK, PMK, MVD, and IDI nucleic acids. The MVA pathway nucleic acids may be heterologous nucleic acids or duplicate copies of endogenous nucleic acids. In some embodiments, the amount of one or more MVA pathway polypeptides is increased by replacing the endogenous promoters or regulatory regions for the MVA pathway nucleic acids with other promoters and/or regulatory regions that result in greater transcription of the MVA pathway nucleic acids. In some embodiments, the cells contain both a heterologous nucleic acid encoding an isoprene synthase polypeptide (e.g., a plant isoprene synthase nucleic acid) and a duplicate copy of an endogenous nucleic acid encoding an isoprene synthase polypeptide.
In some embodiments, at least a portion of the cells maintain the heterologous isoprene synthase, DXS, IDI, and/or MVA pathway nucleic acid for at least about 5, 10, 20, 50, 75, 100, 200, 300, or more cell divisions in a continuous culture (such as a continuous culture without dilution). In some embodiments of any of the aspects of the invention, the nucleic acid comprising the heterologous or duplicate copy of an endogenous isoprene synthase, DXS, IDI, and/or MVA pathway nucleic acid also comprises a selective marker, such as a kanamycin, ampicillin, carbenicillin, gentamicin, hygromycin, phleomycin, bleomycin, neomycin, or chloramphenicol antibiotic resistance nucleic acid.
I. Exemplary Polypeptides and Nucleic Acids
Various isoprene synthase, DXS, IDI, and/or MVA pathway polypeptides and nucleic acids can be used in the compositions and methods of the invention.
As used herein, “polypeptides” includes polypeptides, proteins, peptides, fragments of polypeptides, and fusion polypeptides that include part or all of a first polypeptide (e.g., an isoprene synthase, DXS, IDI, or MVA pathway polypeptide) and part or all of a second polypeptide (e.g., a peptide that facilitates purification or detection of the fusion polypeptide, such as a His-tag). In some embodiments, the fusion polypeptide has an activity of two or more MVA pathway polypeptides (such as AA-CoA thiolase and HMG-CoA reductase polypeptides). In some embodiments, the polypeptide is a naturally-occurring polypeptide (such as the polypeptide encoded by an Enterococcus faecalis mvaE nucleic acid) that has an activity of two or more MVA pathway polypeptides.
In various embodiments, a polypeptide has at least or about 50, 100, 150, 175, 200, 250, 300, 350, 400, or more amino acids. In some embodiments, the polypeptide fragment contains at least or about 25, 50, 75, 100, 150, 200, 300, or more contiguous amino acids from a full-length polypeptide and has at least or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% of an activity of a corresponding full-length polypeptide. In particular embodiments the polypeptide includes a segment of or the entire amino acid sequence of any naturally-occurring isoprene synthase, DXS, IDI, or MVA pathway polypeptide. In some embodiments, the polypeptide has one or more mutations compared to the sequence of a wild-type (i.e., a sequence occurring in nature) isoprene synthase, DXS, IDI, or MVA pathway polypeptide.
In some embodiments, the polypeptide is an isolated polypeptide. As used herein, an “isolated polypeptide” is not part of a library of polypeptides, such as a library of 2, 5, 10, 20, 50 or more different polypeptides and is separated from at least one component with which it occurs in nature. An isolated polypeptide can be obtained, for example, by expression of a recombinant nucleic acid encoding the polypeptide.
In some embodiments, the polypeptide is a heterologous polypeptide. By “heterologous polypeptide” is meant a polypeptide whose amino acid sequence is not identical to that of another polypeptide naturally expressed in the same host cell.
As used herein, a “nucleic acid” refers to two or more deoxyribonucleotides and/or ribonucleotides in either single or double-stranded form. In some embodiments, the nucleic acid is a recombinant nucleic acid. By “recombinant nucleic acid” means a nucleic acid of interest that is free of one or more nucleic acids (e.g., genes), which in the genome occurring in nature of the organism from which the nucleic acid of interest is derived, flank the nucleic acid of interest. The term therefore includes, for example, a recombinant DNA which is incorporated into a vector, into an autonomously replicating plasmid or virus, or into the genomic DNA of a prokaryote or eukaryote, or which exists as a separate molecule (e.g., a cDNA, a genomic DNA fragment, or a cDNA fragment produced by PCR or restriction endonuclease digestion) independent of other sequences.
In various embodiments, the nucleic acid is a recombinant nucleic acid. For instance, in some embodiments, an isoprene synthase, DXS, IDI, or MVA pathway nucleic acid is operably linked to another nucleic acid encoding all or a portion of another polypeptide such that the recombinant nucleic acid encodes a fusion polypeptide that includes an isoprene synthase, DXS, IDI, or MVA pathway polypeptide and all or part of another polypeptide (e.g., a peptide that facilitates purification or detection of the fusion polypeptide, such as a His-tag). In some embodiments, part or all of a recombinant nucleic acid is chemically synthesized. In some embodiments, the nucleic acid is a heterologous nucleic acid. By “heterologous nucleic acid” is meant a nucleic acid whose nucleic acid sequence is not identical to that of another nucleic acid naturally found in the same host cell.
In particular embodiments the nucleic acid includes a segment of or the entire nucleic acid sequence of any naturally-occurring isoprene synthase, DXS, IDI, or MVA pathway nucleic acid. In some embodiments, the nucleic acid includes at least or about 50, 100, 150, 200, 300, 400, 500, 600, 700, 800, or more contiguous nucleotides from a naturally-occurring isoprene synthase nucleic acid DXS, IDI, or MVA pathway nucleic acid. In some embodiments, the nucleic acid has one or more mutations compared to the sequence of a wild-type (i.e., a sequence occurring in nature) isoprene synthase, DXS, IDI, or MVA pathway nucleic acid. In some embodiments, the nucleic acid has one or more mutations (e.g., a silent mutation) that increase the transcription or translation of isoprene synthase, DXS, IDI, or MVA pathway nucleic acid. In some embodiments, the nucleic acid is a degenerate variant of any nucleic acid encoding an isoprene synthase, DXS, IDI, or MVA pathway polypeptide.
“Codon degeneracy” refers to divergence in the genetic code permitting variation of the nucleotide sequence without affecting the amino acid sequence of an encoded polypeptide. The skilled artisan is well aware of the “codon-bias” exhibited by a specific host cell in usage of nucleotide codons to specify a given amino acid. Therefore, when synthesizing a nucleic acid for improved expression in a host cell, it is desirable in some embodiments to design the nucleic acid such that its frequency of codon usage approaches the frequency of preferred codon usage of the host cell.
The accession numbers of exemplary isoprene synthase, DXS, IDI, and/or MVA pathway polypeptides and nucleic acids are listed in Appendix 1 of U.S. Application No. 61/013,574, herein incorporated by reference in its entirety, particularly with respect to the amino acid and nucleic acid sequences of isoprene synthase, DXS, IDI, and/or MVA pathway polypeptides and nucleic acids). The Kegg database also contains the amino acid and nucleic acid sequences of numerous exemplary isoprene synthase, DXS, IDI, and/or MVA pathway polypeptides and nucleic acids (See, e.g., the world-wide web at “genome.jp/kegg/pathway/map/map00100.html” and the sequences therein, which are each hereby incorporated by reference in their entireties, particularly with respect to the amino acid and nucleic acid sequences of isoprene synthase, DXS, IDI, and/or MVA pathway polypeptides and nucleic acids). In some embodiments, one or more of the isoprene synthase, DXS, IDI, and/or MVA pathway polypeptides and/or nucleic acids have a sequence identical to a sequence publicly available on Dec. 12, 2007, such as any of the sequences that correspond to any of the accession numbers in Appendix 1 of U.S. Application No. 61/013,574, or any of the sequences present in the Kegg database as of the date of this filing. Additional exemplary isoprene synthase, DXS, IDI, and/or MVA pathway polypeptides and nucleic acids are described further below.
Exemplary Isoprene Synthase Polypeptides and Nucleic Acids
As noted above, isoprene synthase polypeptides convert dimethylallyl diphosphate (DMAPP) into isoprene. Exemplary isoprene synthase polypeptides include polypeptides, fragments of polypeptides, peptides, and fusions polypeptides that have at least one activity of an isoprene synthase polypeptide. Standard methods can be used to determine whether a polypeptide has isoprene synthase polypeptide activity by measuring the ability of the polypeptide to convert DMAPP into isoprene in vitro, in a cell extract, or in vivo. Isoprene synthase polypeptide activity in the cell extract can be measured, for example, as described in Silver et al., J. Biol. Chem. 270:13010-13016, 1995 and references therein, which are each hereby incorporated by reference in their entireties, particularly with respect to assays for isoprene synthase polypeptide activity. DMAPP (Sigma) is evaporated to dryness under a stream of nitrogen and rehydrated to a concentration of 100 mM in 100 mM potassium phosphate buffer pH 8.2 and stored at −20° C. To perform the assay, a solution of 5 μl of 1M MgCl2, 1 mM (250 μg/ml) DMAPP, 65 μl of Plant Extract Buffer (PEB) (50 mM Tris-HCl, pH 8.0, 20 mM MgCl2, 5% glycerol, and 2 mM DTT) is added to 25 μl of cell extract in a 20 ml Headspace vial with a metal screw cap and teflon coated silicon septum (Agilent Technologies) and cultured at 37° C. for 15 minutes with shaking. The reaction is quenched by adding 200 μl of 250 mM EDTA or by heat inactivation, and isoprene is quantified by GC/MS.
Exemplary isoprene synthase nucleic acids include nucleic acids that encode a polypeptide, fragment of a polypeptide, peptide, or fusion polypeptide that has at least one activity of an isoprene synthase polypeptide. Exemplary isoprene synthase polypeptides and nucleic acids include naturally-occurring polypeptides and nucleic acids from any of the source organisms described herein as well as mutant polypeptides and nucleic acids derived from any of the source organisms described herein.
In some embodiments, the isoprene synthase polypeptide or nucleic acid is from the family Fabaceae, the family Salicaceae, or the family Fagaceae. In some embodiments, the isoprene synthase polypeptide or nucleic acid is a naturally-occurring polypeptide or nucleic acid from Pueraria montana (kudzu) (Sharkey et al., Plant Physiology 137: 700-712, 2005), poplar (such as Populus alba x tremula CAC35696) Miller et al., Planta 213: 483-487, 2001) aspen (such as Populus tremuloides) Silver et al., JBC 270(22): 13010-1316, 1995), or English Oak (Quercus robur) (Zimmer et al., WO 98/02550), which are each hereby incorporated by reference in their entireties, particularly with respect to isoprene synthase nucleic acids and the expression of isoprene synthase polypeptides. Suitable isoprene synthases include, but are not limited to, those identified by GenBank Accession Nos. AY341431, AY316691, AY279379, AJ457070, and AY182241, which are each hereby incorporated by reference in their entireties, particularly with respect to sequences of isoprene synthase nucleic acids and polypeptides. In some embodiments, the isoprene synthase polypeptide or nucleic acid is not a naturally-occurring polypeptide or nucleic acid from Quercus robur (i.e., the isoprene synthase polypeptide or nucleic acid is an isoprene synthase polypeptide or nucleic acid other than a naturally-occurring polypeptide or nucleic acid from Quercus robur). In some embodiments, the isoprene synthase nucleic acid or polypeptide is not a naturally-occurring polypeptide or nucleic acid from poplar (such as Populus alba x tremula CAC35696).
Exemplary DXS Polypeptides and Nucleic Acids
As noted above, 1-deoxy-D-xylulose-5-phosphate synthase (DXS) polypeptides convert pyruvate and D-glyceraldehyde-3-phosphate into 1-deoxy-D-xylulose-5-phosphate. Exemplary DXS polypeptides include polypeptides, fragments of polypeptides, peptides, and fusions polypeptides that have at least one activity of a DXS polypeptide. Standard methods (such as those described herein) can be used to determine whether a polypeptide has DXS polypeptide activity by measuring the ability of the polypeptide to convert pyruvate and D-glyceraldehyde-3-phosphate into 1-deoxy-D-xylulose-5-phosphate in vitro, in a cell extract, or in vivo. Exemplary DXS nucleic acids include nucleic acids that encode a polypeptide, fragment of a polypeptide, peptide, or fusion polypeptide that has at least one activity of a DXS polypeptide. Exemplary DXS polypeptides and nucleic acids include naturally-occurring polypeptides and nucleic acids from any of the source organisms described herein as well as mutant polypeptides and nucleic acids derived from any of the source organisms described herein.
Exemplary IDI Polypeptides and Nucleic Acids
Isopentenyl diphosphate isomerase polypeptides (isopentenyl-diphosphate delta-isomerase or IDI) catalzyes the interconversion of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) (e.g., converting IPP into DMAPP and/or converting DMAPP into IPP). Exemplary IDI polypeptides include polypeptides, fragments of polypeptides, peptides, and fusions polypeptides that have at least one activity of an IDI polypeptide. Standard methods (such as those described herein) can be used to determine whether a polypeptide has IDI polypeptide activity by measuring the ability of the polypeptide to interconvert IPP and DMAPP in vitro, in a cell extract, or in vivo. Exemplary IDI nucleic acids include nucleic acids that encode a polypeptide, fragment of a polypeptide, peptide, or fusion polypeptide that has at least one activity of an IDI polypeptide. Exemplary IDI polypeptides and nucleic acids include naturally-occurring polypeptides and nucleic acids from any of the source organisms described herein as well as mutant polypeptides and nucleic acids derived from any of the source organisms described herein.
Exemplary MVA Pathway Polypeptides and Nucleic Acids
Exemplary MVA pathway polypeptides include acetyl-CoA acetyltransferase (AA-CoA thiolase) polypeptides, 3-hydroxy-3-methylglutaryl-CoA synthase (HMG-CoA synthase) polypeptides, 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase) polypeptides, mevalonate kinase (MVK) polypeptides, phosphomevalonate kinase (PMK) polypeptides, diphosphomevalonte decarboxylase (MVD) polypeptides, IDI polypeptides, and polypeptides (e.g., fusion polypeptides) having an activity of two or more MVA pathway polypeptides. In particular, MVA pathway polypeptides include polypeptides, fragments of polypeptides, peptides, and fusions polypeptides that have at least one activity of an MVA pathway polypeptide. Exemplary MVA pathway nucleic acids include nucleic acids that encode a polypeptide, fragment of a polypeptide, peptide, or fusion polypeptide that has at least one activity of an MVA pathway polypeptide. Exemplary MVA pathway polypeptides and nucleic acids include naturally-occurring polypeptides and nucleic acids from any of the source organisms described herein as well as mutant polypeptides and nucleic acids derived from any of the source organisms described herein.
In particular, acetyl-CoA acetyltransferase polypeptides (AA-CoA thiolase or AACT) convert two molecules of acetyl-CoA into acetoacetyl-CoA. Standard methods (such as those described herein) can be used to determine whether a polypeptide has AA-CoA thiolase polypeptide activity by measuring the ability of the polypeptide to convert two molecules of acetyl-CoA into acetoacetyl-CoA in vitro, in a cell extract, or in vivo.
3-hydroxy-3-methylglutaryl-CoA synthase (HMG-CoA synthase or HMGS) polypeptides convert acetoacetyl-CoA into 3-hydroxy-3-methylglutaryl-CoA. Standard methods (such as those described herein) can be used to determine whether a polypeptide has HMG-CoA synthase polypeptide activity by measuring the ability of the polypeptide to convert acetoacetyl-CoA into 3-hydroxy-3-methylglutaryl-CoA in vitro, in a cell extract, or in vivo.
3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase or HMGR) polypeptides convert 3-hydroxy-3-methylglutaryl-CoA into mevalonate. Standard methods (such as those described herein) can be used to determine whether a polypeptide has HMG-CoA reductase polypeptide activity by measuring the ability of the polypeptide to convert 3-hydroxy-3-methylglutaryl-CoA into mevalonate in vitro, in a cell extract, or in vivo.
Mevalonate kinase (MVK) polypeptides phosphorylates mevalonate to form mevalonate-5-phosphate. Standard methods (such as those described herein) can be used to determine whether a polypeptide has MVK polypeptide activity by measuring the ability of the polypeptide to convert mevalonate into mevalonate-5-phosphate in vitro, in a cell extract, or in vivo.
Phosphomevalonate kinase (PMK) polypeptides phosphorylates mevalonate-5-phosphate to form mevalonate-5-diphosphate. Standard methods (such as those described herein) can be used to determine whether a polypeptide has PMK polypeptide activity by measuring the ability of the polypeptide to convert mevalonate-5-phosphate into mevalonate-5-diphosphate in vitro, in a cell extract, or in vivo.
Diphosphomevalonte decarboxylase (MVD or DPMDC) polypeptides convert mevalonate-5-diphosphate into isopentenyl diphosphate polypeptides (IPP). Standard methods (such as those described herein) can be used to determine whether a polypeptide has MVD polypeptide activity by measuring the ability of the polypeptide to convert mevalonate-5-diphosphate into IPP in vitro, in a cell extract, or in vivo.
Exemplary Methods for Isolating Nucleic Acids
Isoprene synthase, DXS, IDI, and/or MVA pathway nucleic acids can be isolated using standard methods. Methods of obtaining desired nucleic acids from a source organism of interest (such as a bacterial genome) are common and well known in the art of molecular biology (see, for example, WO 2004/033646 and references cited therein, which are each hereby incorporated by reference in their entireties, particularly with respect to the isolation of nucleic acids of interest). For example, if the sequence of the nucleic acid is known (such as any of the known nucleic acids described herein), suitable genomic libraries may be created by restriction endonuclease digestion and may be screened with probes complementary to the desired nucleic acid sequence. Once the sequence is isolated, the DNA may be amplified using standard primer directed amplification methods such as polymerase chain reaction (PCR) (U.S. Pat. No. 4,683,202, which is incorporated by reference in its entirety, particularly with respect to PCR methods) to obtain amounts of DNA suitable for transformation using appropriate vectors. Alternatively, isoprene synthase, DXS, IDI, and/or MVA pathway nucleic acids (such as any isoprene synthase, DXS, IDI, and/or MVA pathway nucleic acids with a known nucleic acid sequence) can be chemically synthesized using standard methods.
Additional isoprene synthase, DXS, IDI, or MVA pathway polypeptides and nucleic acids that may be suitable for use in the compositions and methods described herein can be identified using standard methods. For example, cosmid libraries of the chromosomal DNA of organisms known to produce isoprene naturally can be constructed in organisms such as E. coli, and then screened for isoprene production. Additional methods for obtaining isoprene synthase, DXS, IDI, and/or MVA pathway nucleic acids include screening a metagenomic library by assay (such as the headspace assay described herein) or by PCR using primers directed against nucleotides encoding for a length of conserved amino acids (for example, at least 3 conserved amino acids). Conserved amino acids can be identified by aligning amino acid sequences of known isoprene synthase, DXS, IDI, and/or MVA pathway polypeptides. Conserved amino acids for isoprene synthase polypeptides can be identified based on aligned sequences of known isoprene synthase polypeptides. An organism found to produce isoprene naturally can be subjected to standard protein purification methods (which are well known in the art) and the resulting purified polypeptide can be sequenced using standard methods. Other methods are found in the literature (See, e.g., Julsing et al., Applied. Microbiol. Biotechnol. 75: 1377-84, 2007; and Withers et al., Appl Environ Microbiol. 73:6277-83, 2007, which are each hereby incorporated by reference in their entireties, particularly with respect to identification of nucleic acids involved in the synthesis of isoprene).
Additionally, standard sequence alignment and/or structure prediction programs can be used to identify additional DXS, IDI, or MVA pathway polypeptides and nucleic acids based on the similarity of their primary and/or predicted polypeptide secondary structure with that of known DXS, IDI, or MVA pathway polypeptides and nucleic acids. Standard databases such as the swissprot-trembl database (world-wide web at “expasy.org”, Swiss Institute of Bioinformatics Swiss-Prot group CMU—1 rue Michel Servet CH-1211 Geneva 4, Switzerland) can also be used to identify isoprene synthase, DXS, IDI, or MVA pathway polypeptides and nucleic acids. The secondary and/or tertiary structure of an isoprene synthase, DXS, IDI, or MVA pathway polypeptide can be predicted using the default settings of standard structure prediction programs, such as PredictProtein (Rost et al., The PredictProtein Server. Nucleic Acids Research 32(Web Server issue):W321-W326, 2004). Alternatively, the actual secondary and/or tertiary structure of an isoprene synthase, DXS, IDI, or MVA pathway polypeptide can be determined using standard methods. Additional isoprene synthase, DXS, IDI, or MVA pathway nucleic acids can also be identified by hybridization to probes generated from known isoprene synthase, DXS, IDI, or MVA pathway nucleic acids.
Exemplary Promoters and Vectors
Any of the isoprene synthase, DXS, IDI, or MVA pathway nucleic acid described herein can be included in one or more vectors. Accordingly, the invention also features vectors with one more nucleic acids encoding any of the isoprene synthase, DXS, IDI, or MVA pathway polypeptides that are described herein. As used herein, a “vector” means a construct that is capable of delivering, and desirably expressing one or more nucleic acids of interest in a host cell. Examples of vectors include, but are not limited to, plasmids, viral vectors, DNA or RNA expression vectors, cosmids, and phage vectors. In some embodiments, the vector contains a nucleic acid under the control of an expression control sequence.
As used herein, an “expression control sequence” means a nucleic acid sequence that directs transcription of a nucleic acid of interest. An expression control sequence can be a promoter, such as a constitutive or an inducible promoter, or an enhancer. An “inducible promoter” is a promoter that is active under environmental or developmental regulation. The expression control sequence is operably linked to the nucleic acid segment to be transcribed.
In some embodiments, the vector contains a selective marker. The term “selective marker” refers to a nucleic acid capable of expression in a host cell that allows for ease of selection of those host cells containing an introduced nucleic acid or vector. Examples of selectable markers include, but are not limited to, antibiotic resistance nucleic acids (e.g., kanamycin, ampicillin, carbenicillin, gentamicin, hygromycin, phleomycin, bleomycin, neomycin, or chloramphenicol) and/or nucleic acids that confer a metabolic advantage, such as a nutritional advantage on the host cell. In some embodiments, an isoprene synthase, DXS, IDI, or MVA pathway nucleic acid integrates into a chromosome of the cells without a selective marker.
Suitable vectors are those that are compatible with the host cell employed. Suitable vectors can be derived, for example, from a bacterium, a virus (such as bacteriophage T7 or a M-13 derived phage), a cosmid, a yeast, or a plant. Protocols for obtaining and using such vectors are known in the art (See, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor, 1989, which is hereby incorporated by reference in its entirety, particularly with respect to the use of vectors).
Promoters are well known in the art. Any promoter that functions in the host cell can be used for expression of an isoprene synthase, DXS, IDI, or MVA pathway nucleic acid in the host cell. Initiation control regions or promoters, which are useful to drive expression of isoprene synthase, DXS, IDI, or MVA pathway nucleic acids in various host cells are numerous and familiar to those skilled in the art (see, for example, WO 2004/033646 and references cited therein, which are each hereby incorporated by reference in their entireties, particularly with respect to vectors for the expression of nucleic acids of interest). Virtually any promoter capable of driving these nucleic acids is suitable for the present invention including, but not limited to lac, trp, λPL, λPR, T7, tac, and trc (useful for expression in E. coli).
In some embodiments, a glucose isomerase promoter is used (see, for example, U.S. Pat. No. 7,132,527 and references cited therein, which are each hereby incorporated by reference in their entireties, particularly with respect promoters and plasmid systems for expressing polypeptides of interest). Reported glucose isomerase promoter mutants can be used to vary the level of expression of the polypeptide encoded by a nucleic acid operably linked to the glucose isomerase promoter (U.S. Pat. No. 7,132,527). In various embodiments, the glucose isomerase promoter is contained in a low, medium, or high copy plasmid (U.S. Pat. No. 7,132,527).
In various embodiments, an isoprene synthase, DXS, IDI, and/or MVA pathway nucleic acid is contained in a low copy plasmid (e.g., a plasmid that is maintained at about 1 to about 4 copies per cell), medium copy plasmid (e.g., a plasmid that is maintained at about 10 to about 15 copies per cell), or high copy plasmid (e.g., a plasmid that is maintained at about 50 or more copies per cell). In some embodiments, the heterologous or extra endogenous isoprene synthase, DXS, IDI, or MVA pathway nucleic acid is operably linked to a T7 promoter. In some embodiments, the heterologous or extra endogenous isoprene synthase, DXS, IDI, or MVA pathway nucleic acid operably linked to a T7 promoter is contained in a medium or high copy plasmid. In some embodiments, the heterologous or extra endogenous isoprene synthase, DXS, IDI, or MVA pathway nucleic acid is operably linked to a Trc promoter. In some embodiments, the heterologous or extra endogenous isoprene synthase, DXS, IDI, or MVA pathway nucleic acid operably linked to a Trc promoter is contained in a medium or high copy plasmid. In some embodiments, the heterologous or extra endogenous isoprene synthase, DXS, IDI, or MVA pathway nucleic acid is operably linked to a Lac promoter. In some embodiments, the heterologous or extra endogenous isoprene synthase, DXS, IDI, or MVA pathway nucleic acid operably linked to a Lac promoter is contained in a low copy plasmid. In some embodiments, the heterologous or extra endogenous isoprene synthase, DXS, IDI, or MVA pathway nucleic acid is operably linked to an endogenous promoter, such as an endogenous Escherichia, Panteoa, Bacillus, Yarrowia, Streptomyces, or Trichoderma promoter or an endogenous alkaline serine protease, isoprene synthase, DXS, IDI, or MVA pathway promoter. In some embodiments, the heterologous or extra endogenous isoprene synthase, DXS, IDI, or MVA pathway nucleic acid operably linked to an endogenous promoter is contained in a high copy plasmid. In some embodiments, the vector is a replicating plasmid that does not integrate into a chromosome in the cells. In some embodiments part or all of the vector integrates into a chromosome in the cells.
In some embodiments, the expression vector also includes a termination sequence. Termination control regions may also be derived from various genes native to the host cell. In some embodiments, the termination sequence and the promoter sequence are derived from the same source. In another embodiment, the termination sequence is endogenous to the host cell.
In some embodiments, the promoter, coding, region, and terminator all originate from the isoprene synthase, DXS, IDI, or MVA pathway nucleic acid to be expressed. In some embodiments, the coding region for an isoprene synthase, DXS, IDI, or MVA pathway nucleic acid is inserted into a general-purpose expression vector such that it is under the transcriptional control of the expression construct promoter and terminator sequences. In some embodiments, genes or part thereof are inserted downstream of the strong cbh1 promoter.
An isoprene synthase, DXS, IDI, or MVA pathway nucleic acid can be incorporated into a vector, such as an expression vector, using standard techniques (Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor, 1982, which is hereby incorporated by reference in its entirety, particularly with respect to the screening of appropriate DNA sequences and the construction of vectors). Methods used to ligate the DNA construct comprising a nucleic acid of interest (such as an isoprene synthase, DXS, IDI, or MVA pathway nucleic acid), a promoter, a terminator, and other sequences and to insert them into a suitable vector are well known in the art. For example, restriction enzymes can be used to cleave the isoprene synthase, DXS, IDI, or MVA pathway nucleic acid and the vector. Then, the compatible ends of the cleaved isoprene synthase, DXS, IDI, or MVA pathway nucleic acid and the cleaved vector can be ligated. Linking is generally accomplished by ligation at convenient restriction sites. If such sites do not exist, the synthetic oligonucleotide linkers are used in accordance with conventional practice (see, Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor, 1989, and Bennett and Lasure, More Gene Manipulations in Fungi, Academic Press, San Diego, pp 70-76, 1991, which are each hereby incorporated by reference in their entireties, particularly with respect to oligonucleotide linkers). Additionally, vectors can be constructed using known recombination techniques (e.g., Invitrogen Life Technologies, Gateway Technology).
In some embodiments, it may be desirable to over-express isoprene synthase, DXS, IDI, or MVA pathway nucleic acids at levels far higher than currently found in naturally-occurring cells. This result may be accomplished by the selective cloning of the nucleic acids encoding those polypeptides into multicopy plasmids or placing those nucleic acids under a strong inducible or constitutive promoter. Methods for over-expressing desired polypeptides are common and well known in the art of molecular biology and examples may be found in Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor, 1989, which is hereby incorporated by reference in its entirety, particularly with respect to cloning techniques.
The following resources include descriptions of additional general methodology useful in accordance with the invention: Kreigler, Gene Transfer and Expression; A Laboratory Manual, 1990; and Ausubel et al., Eds. Current Protocols in Molecular Biology, 1994, which are each hereby incorporated by reference in their entireties, particularly with respect to molecular biology and cloning techniques.
Exemplary Source Organisms
Isoprene synthase, DXS, IDI, or MVA pathway nucleic acids (and their encoded polypeptides) can be obtained from any organism that naturally contains isoprene synthase, DXS, IDI, and/or MVA pathway nucleic acids. As noted above, isoprene is formed naturally by a variety of organisms, such as bacteria, yeast, plants, and animals. Organisms contain the MVA pathway, DXP pathway, or both the MVA and DXP pathways for producing isoprene (FIG. 15). Thus, DXS nucleic acids can be obtained, e.g., from any organism that contains the DXP pathway or contains both the MVA and DXP pathways. IDI and isoprene synthase nucleic acids can be obtained, e.g., from any organism that contains the MVA pathway, DXP pathway, or both the MVA and DXP pathways. MVA pathway nucleic acids can be obtained, e.g., from any organism that contains the MVA pathway or contains both the MVA and DXP pathways.
In some embodiments, the nucleic acid sequence of the isoprene synthase, DXS, IDI, or MVA pathway nucleic is identical to the sequence of a nucleic acid that is produced by any of the following organisms in nature. In some embodiments, the amino acid sequence of the isoprene synthase, DXS, IDI, or MVA pathway polypeptide is identical to the sequence of a polypeptide that is produced by any of the following organisms in nature. In some embodiments, the isoprene synthase, DXS, IDI, or MVA pathway nucleic acid or polypeptide is a mutant nucleic acid or polypeptide derived from any of the organisms described herein. As used herein, “derived from” refers to the source of the nucleic acid or polypeptide into which one or more mutations is introduced. For example, a polypeptide that is “derived from a plant polypeptide” refers to polypeptide of interest that results from introducing one or more mutations into the sequence of a wild-type (i.e., a sequence occurring in nature) plant polypeptide.
In some embodiments, the source organism is a bacterium, such as strains of Escherichia (e.g., E. coli), or strains of Bacillus (e.g., B. subtilis).
As used herein, “the genus Escherichia” includes all species within the genus “Escherichia,” as known to those of skill in the art, including but not limited to E. coli, E. adecarboxylata, E. albertii, E. blattae, E. fergusonii, E. hermannii, E. senegalensis, and E. vulneris. The genus “Escherichia” is defined as Gram-negative, non-spore forming, facultatively anaerobic, rod-shaped bacteria are classified as members of the Family Enterobacteriaceae, Order Enterobacteriales, Class Gamma Proteobacteria.
As used herein, “the genus Bacillus” includes all species within the genus “Bacillus,” as known to those of skill in the art, including but not limited to B. subtilis, B. licheniformis, B. lentus, B. brevis, B. stearothermophilus, B. alkalophilus, B. amyloliquefaciens, B. clausii, B. halodurans, B. megaterium, B. coagulans, B. circulans, B. lautus, and B. thuringiensis. It is recognized that the genus Bacillus continues to undergo taxonomical reorganization. Thus, it is intended that the genus include species that have been reclassified, including but not limited to such organisms as B. stearothermophilus, which is now named “Geobacillus stearothermophilus.” The production of resistant endospores in the presence of oxygen is considered the defining feature of the genus Bacillus, although this characteristic also applies to the recently named Alicyclobacillus, Amphibacillus, Aneurinibacillus, Anoxybacillus, Brevibacillus, Filobacillus, Gracilibacillus, Halobacillus, Paenibacillus, Salibacillus, Thermobacillus, Ureibacillus, and Virgibacillus. 
Exemplary Host Cells
A variety of host cells can be used to express isoprene synthase, DXS, IDI, and/or MVA pathway polypeptides and to produce isoprene in the methods of the claimed invention. Exemplary host cells include cells from any of the organisms listed in the prior section under the heading “Exemplary Source Organisms.” The host cell may be a cell that naturally produces isoprene or a cell that does not naturally produce isoprene. In some embodiments, the host cell naturally produces isoprene using the DXP pathway, and an isoprene synthase, DXS, and/or IDI nucleic acid is added to enhance production of isoprene using this pathway. In some embodiments, the host cell naturally produces isoprene using the MVA pathway, and an isoprene synthase and/or one or more MVA pathway nucleic acids are added to enhance production of isoprene using this pathway. In some embodiments, the host cell naturally produces isoprene using the DXP pathway and one or more MVA pathway nucleic acids are added to produce isoprene using part or all of the MVA pathway as well as the DXP pathway. In some embodiments, the host cell naturally produces isoprene using both the DXP and MVA pathways and one or more isoprene synthase, DXS, IDI, or MVA pathway nucleic acids are added to enhance production of isoprene by one or both of these pathways.
Exemplary Transformation Methods
Isoprene synthase, DXS, IDI, and/or MVA pathway nucleic acids or vectors containing them can be inserted into a host cell (e.g., a bacterial cell) using standard techniques for expression of the encoded isoprene synthase, DXS, IDI, and/or MVA pathway polypeptide. Introduction of a DNA construct or vector into a host cell can be performed using techniques such as transformation, electroporation, nuclear microinjection, transduction, transfection (e.g., lipofection mediated or DEAE-Dextrin mediated transfection or transfection using a recombinant phage virus), incubation with calcium phosphate DNA precipitate, high velocity bombardment with DNA-coated microprojectiles, and protoplast fusion. General transformation techniques are known in the art (see, e.g., Current Protocols in Molecular Biology (F. M. Ausubel et al. (eds) Chapter 9, 1987; Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor, 1989; and Campbell et al., Curr Genet, 16:53-56, 1989, which are each hereby incorporated by reference in their entireties, particularly with respect to transformation methods). The introduced nucleic acids may be integrated into chromosomal DNA or maintained as extrachromosomal replicating sequences.
Exemplary Cell Culture Media
The invention also includes a cell or a population of cells in culture that produce isoprene. By “cells in culture” is meant two or more cells in a solution (e.g., a cell medium) that allows the cells to undergo one or more cell divisions. “Cells in culture” do not include plant cells that are part of a living, multicellular plant containing cells that have differentiated into plant tissues. In various embodiments, the cell culture includes at least or about 10, 20, 50, 100, 200, 500, 1,000, 5,000, 10,000 or more cells.
Any carbon source can be used to cultivate the host cells. The term “carbon source” refers to one or more carbon-containing compounds capable of being metabolized by a host cell or organism. For example, the cell medium used to cultivate the host cells may include any carbon source suitable for maintaining the viability or growing the host cells.
In some embodiments, the carbon source is a carbohydrate (such as monosaccharide, disaccharide, oligosaccharide, or polysaccharides), invert sugar (e.g., enzymatically treated sucrose syrup), glycerol, glycerine (e.g., a glycerine byproduct of a biodiesel or soap-making process), dihydroxyacetone, one-carbon source, fatty acid (e.g., a saturated fatty acid, unsaturated fatty acid, or polyunsaturated fatty acid), lipid, phospholipid, glycerolipid, monoglyceride, diglyceride, triglyceride, polypeptide (e.g., a microbial or plant protein or peptide), renewable carbon source (e.g., a biomass carbon source such as a hydrolyzed biomass carbon source; beet sugar or cane sugar molasses), yeast extract, component from a yeast extract, polymer, acid, alcohol, aldehyde, ketone, amino acid, succinate, lactate, acetate, ethanol, or any combination of two or more of the foregoing. In some embodiments, the carbon source is a product of photosynthesis, including, but not limited to, glucose.
Exemplary monosaccharides include glucose and fructose; exemplary oligosaccharides include lactose and sucrose, and exemplary polysaccharides include starch and cellulose. Exemplary carbohydrates include C6 sugars (e.g., fructose, mannose, galactose, or glucose) and C5 sugars (e.g., xylose or arabinose). In some embodiments, the cell medium includes a carbohydrate as well as a carbon source other than a carbohydrate (e.g., glycerol, glycerine, dihydroxyacetone, one-carbon source, fatty acid, lipid, phospholipid, glycerolipid, monoglyceride, diglyceride, triglyceride, renewable carbon source, or a component from a yeast extract). In some embodiments, the cell medium includes a carbohydrate as well as a polypeptide (e.g., a microbial or plant protein or peptide). In some embodiments, the microbial polypeptide is a polypeptide from yeast or bacteria. In some embodiments, the plant polypeptide is a polypeptide from soy, corn, canola, jatropha, palm, peanut, sunflower, coconut, mustard, rapeseed, cottonseed, palm kernel, olive, safflower, sesame, or linseed.
In some embodiments, the concentration of the carbohydrate is at least or about 5 grams per liter of broth (g/L, wherein the volume of broth includes both the volume of the cell medium and the volume of the cells), such as at least or about 10, 15, 20, 30, 40, 50, 60, 80, 100, 150, 200, 300, 400, or more g/L. In some embodiments, the concentration of the carbohydrate is between about 50 and about 400 g/L, such as between about 100 and about 360 g/L, between about 120 and about 360 g/L, or between about 200 and about 300 g/L. In some embodiments, this concentration of carbohydrate includes the total amount of carbohydrate that is added before and/or during the culturing of the host cells.
Exemplary lipids are any substance containing one or more fatty acids that are C4 and above fatty acids that are saturated, unsaturated, or branched.
Exemplary fatty acids include compounds of the formula R—COOH, where “R” is a hydrocarbon. Exemplary unsaturated fatty acids include compounds where “R” includes at least one carbon-carbon double bond. Exemplary unsaturated fatty acids include, but are not limited to, oleic acid, vaccenic acid, linoleic acid, palmitelaidic acid, and arachidonic acid. Exemplary polyunsaturated fatty acids include compounds where “R” includes a plurality of carbon-carbon double bonds. Exemplary saturated fatty acids include compounds where “R” is a saturated aliphatic group. In some embodiments, the carbon source includes one or more C12-C22 fatty acids, such as a C12 saturated fatty acid, a C14 saturated fatty acid, a C16 saturated fatty acid, a C18 saturated fatty acid, a C20 saturated fatty acid, or a C22 saturated fatty acid. In an exemplary embodiment, the fatty acid is palmitic acid. In some embodiments, the carbon source is a salt of a fatty acid (e.g., an unsaturated fatty acid), a derivative of a fatty acid (e.g., an unsaturated fatty acid), or a salt of a derivative of fatty acid (e.g., an unsaturated fatty acid). Suitable salts include, but are not limited to, lithium salts, potassium salts, sodium salts, and the like. Di- and triglycerols are fatty acid esters of glycerol.
In some embodiments, the concentration of the lipid, fatty acid, monoglyceride, diglyceride, or triglyceride is at least or about 1 gram per liter of broth (g/L, wherein the volume of broth includes both the volume of the cell medium and the volume of the cells), such as at least or about 5, 10, 15, 20, 30, 40, 50, 60, 80, 100, 150, 200, 300, 400, or more g/L. In some embodiments, the concentration of the lipid, fatty acid, monoglyceride, diglyceride, or triglyceride is between about 10 and about 400 g/L, such as between about 25 and about 300 g/L, between about 60 and about 180 g/L, or between about 75 and about 150 g/L. In some embodiments, the concentration includes the total amount of the lipid, fatty acid, monoglyceride, diglyceride, or triglyceride that is added before and/or during the culturing of the host cells. In some embodiments, the carbon source includes both (i) a lipid, fatty acid, monoglyceride, diglyceride, or triglyceride and (ii) a carbohydrate, such as glucose. In some embodiments, the ratio of the lipid, fatty acid, monoglyceride, diglyceride, or triglyceride to the carbohydrate is about 1:1 on a carbon basis (i.e., one carbon in the lipid, fatty acid, monoglyceride, diglyceride, or triglyceride per carbohydrate carbon). In particular embodiments, the amount of the lipid, fatty acid, monoglyceride, diglyceride, or triglyceride is between about 60 and 180 g/L, and the amount of the carbohydrate is between about 120 and 360 g/L.
Exemplary microbial polypeptide carbon sources include one or more polypeptides from yeast or bacteria. Exemplary plant polypeptide carbon sources include one or more polypeptides from soy, corn, canola, jatropha, palm, peanut, sunflower, coconut, mustard, rapeseed, cottonseed, palm kernel, olive, safflower, sesame, or linseed.
Exemplary renewable carbon sources include cheese whey permeate, cornsteep liquor, sugar beet molasses, barley malt, and components from any of the foregoing. Exemplary renewable carbon sources also include glucose, hexose, pentose and xylose present in biomass, such as corn, switchgrass, sugar cane, cell waste of fermentation processes, and protein by-product from the milling of soy, corn, or wheat. In some embodiments, the biomass carbon source is a lignocellulosic, hemicellulosic, or cellulosic material such as, but are not limited to, a grass, wheat, wheat straw, bagasse, sugar cane bagasse, soft wood pulp, corn, corn cob or husk, corn kernel, fiber from corn kernels, corn stover, switch grass, rice hull product, or a by-product from wet or dry milling of grains (e.g., corn, sorghum, rye, triticate, barley, wheat, and/or distillers grains). Exemplary cellulosic materials include wood, paper and pulp waste, herbaceous plants, and fruit pulp. In some embodiments, the carbon source includes any plant part, such as stems, grains, roots, or tubers. In some embodiments, all or part of any of the following plants are used as a carbon source: corn, wheat, rye, sorghum, triticate, rice, millet, barley, cassaya, legumes, such as beans and peas, potatoes, sweet potatoes, bananas, sugarcane, and/or tapioca. In some embodiments, the carbon source is a biomass hydrolysate, such as a biomass hydrolysate that includes both xylose and glucose or that includes both sucrose and glucose.
In some embodiments, the renewable carbon source (such as biomass) is pretreated before it is added to the cell culture medium. In some embodiments, the pretreatment includes enzymatic pretreatment, chemical pretreatment, or a combination of both enzymatic and chemical pretreatment (see, for example, Farzaneh et al., Bioresource Technology 96 (18): 2014-2018, 2005; U.S. Pat. No. 6,176,176; U.S. Pat. No. 6,106,888; which are each hereby incorporated by reference in their entireties, particularly with respect to the pretreatment of renewable carbon sources). In some embodiments, the renewable carbon source is partially or completely hydrolyzed before it is added to the cell culture medium.
In some embodiments, the renewable carbon source (such as corn stover) undergoes ammonia fiber expansion (AFEX) pretreatment before it is added to the cell culture medium (see, for example, Farzaneh et al., Bioresource Technology 96 (18): 2014-2018, 2005). During AFEX pretreatment, a renewable carbon source is treated with liquid anhydrous ammonia at moderate temperatures (such as about 60 to about 100° C.) and high pressure (such as about 250 to about 300 psi) for about 5 minutes. Then, the pressure is rapidly released. In this process, the combined chemical and physical effects of lignin solubilization, hemicellulose hydrolysis, cellulose decrystallization, and increased surface area enables near complete enzymatic conversion of cellulose and hemicellulose to fermentable sugars. AFEX pretreatment has the advantage that nearly all of the ammonia can be recovered and reused, while the remaining serves as nitrogen source for microbes in downstream processes. Also, a wash stream is not required for AFEX pretreatment. Thus, dry matter recovery following the AFEX treatment is essentially 100%. AFEX is basically a dry-to-dry process. The treated renewable carbon source is stable for long periods and can be fed at very high solid loadings in enzymatic hydrolysis or fermentation processes. Cellulose and hemicellulose are well preserved in the AFEX process, with little or no degradation. There is no need for neutralization prior to the enzymatic hydrolysis of a renewable carbon source that has undergone AFEX pretreatment. Enzymatic hydrolysis of AFEX-treated carbon sources produces clean sugar streams for subsequent fermentation use.
In some embodiments, the concentration of the carbon source (e.g., a renewable carbon source) is equivalent to at least or about 0.1, 0.5, 1, 1.5 2, 3, 4, 5, 10, 15, 20, 30, 40, or 50% glucose (w/v). The equivalent amount of glucose can be determined by using standard HPLC methods with glucose as a reference to measure the amount of glucose generated from the carbon source. In some embodiments, the concentration of the carbon source (e.g., a renewable carbon source) is equivalent to between about 0.1 and about 20% glucose, such as between about 0.1 and about 10% glucose, between about 0.5 and about 10% glucose, between about 1 and about 10% glucose, between about 1 and about 5% glucose, or between about 1 and about 2% glucose.
In some embodiments, the carbon source includes yeast extract or one or more components of yeast extract. In some embodiments, the concentration of yeast extract is at least 1 gram of yeast extract per liter of broth (g/L, wherein the volume of broth includes both the volume of the cell medium and the volume of the cells), such at least or about 5, 10, 15, 20, 30, 40, 50, 60, 80, 100, 150, 200, 300, or more g/L. In some embodiments, the concentration of yeast extract is between about 1 and about 300 g/L, such as between about 1 and about 200 g/L, between about 5 and about 200 g/L, between about 5 and about 100 g/L, or between about 5 and about 60 g/L. In some embodiments, the concentration includes the total amount of yeast extract that is added before and/or during the culturing of the host cells. In some embodiments, the carbon source includes both yeast extract (or one or more components thereof) and another carbon source, such as glucose. In some embodiments, the ratio of yeast extract to the other carbon source is about 1:5, about 1:10, or about 1:20 (w/w).
Additionally the carbon source may also be one-carbon substrates such as carbon dioxide, or methanol. Glycerol production from single carbon sources (e.g., methanol, formaldehyde, or formate) has been reported in methylotrophic yeasts (Yamada et al., Agric. Biol. Chem., 53(2) 541-543, 1989, which is hereby incorporated by reference in its entirety, particularly with respect to carbon sources) and in bacteria (Hunter et. al., Biochemistry, 24, 4148-4155, 1985, which is hereby incorporated by reference in its entirety, particularly with respect to carbon sources). These organisms can assimilate single carbon compounds, ranging in oxidation state from methane to formate, and produce glycerol. The pathway of carbon assimilation can be through ribulose monophosphate, through serine, or through xylulose-momophosphate (Gottschalk, Bacterial Metabolism, Second Edition, Springer-Verlag: New York, 1986, which is hereby incorporated by reference in its entirety, particularly with respect to carbon sources). The ribulose monophosphate pathway involves the condensation of formate with ribulose-5-phosphate to form a six carbon sugar that becomes fructose and eventually the three carbon product glyceraldehyde-3-phosphate Likewise, the serine pathway assimilates the one-carbon compound into the glycolytic pathway via methylenetetrahydrofolate.
In addition to one and two carbon substrates, methylotrophic organisms are also known to utilize a number of other carbon containing compounds such as methylamine, glucosamine and a variety of amino acids for metabolic activity. For example, methylotrophic yeast are known to utilize the carbon from methylamine to form trehalose or glycerol (Bellion et al., Microb. Growth Cl Compd., [Int. Symp.], 7th ed., 415-32. Editors: Murrell et al., Publisher: Intercept, Andover, UK, 1993, which is hereby incorporated by reference in its entirety, particularly with respect to carbon sources). Similarly, various species of Candida metabolize alanine or oleic acid (Sulter et al., Arch. Microbiol. 153(5), 485-9, 1990, which is hereby incorporated by reference in its entirety, particularly with respect to carbon sources).
In some embodiments, cells are cultured in a standard medium containing physiological salts and nutrients (see, e.g., Pourquie, J. et al., Biochemistry and Genetics of Cellulose Degradation, eds. Aubert et al., Academic Press, pp. 71-86, 1988; and Ilmen et al., Appl. Environ. Microbiol. 63:1298-1306, 1997, hereby incorporated by reference, particularly with respect to cell media). Exemplary growth media are common commercially prepared media such as Luria Bertani (LB) broth, Sabouraud Dextrose (SD) broth, or Yeast medium (YM) broth. Other defined or synthetic growth media may also be used, and the appropriate medium for growth of particular host cells are known by someone skilled in the art of microbiology or fermentation science.
In addition to an appropriate carbon source, the cell medium desirably contains suitable minerals, salts, cofactors, buffers, and other components known to those skilled in the art suitable for the growth of the cultures or the enhancement of isoprene production (see, for example, WO 2004/033646 and references cited therein and WO 96/35796 and references cited therein, which are each hereby incorporated by reference in their entireties, particularly with respect cell medias and cell culture conditions). In some embodiments where an isoprene synthase, DXS, IDI, and/or MVA pathway nucleic acid is under the control of an inducible promoter, the inducing agent (e.g., a sugar, metal salt or antimicrobial), is desirably added to the medium at a concentration effective to induce expression of an isoprene synthase, DXS, IDI, and/or MVA pathway polypeptide. In some embodiments, cell medium has an antibiotic (such as kanamycin) that corresponds to the antibiotic resistance nucleic acid (such as a kanamycin resistance nucleic acid) on a vector that has one or more DXS, IDI, or MVA pathway nucleic acids.
Exemplary Production of Isoprene
In some embodiments, the cells are cultured in a culture medium under conditions permitting the production of isoprene by the cells. In some embodiments, the cells in culture produce isoprene at greater than or about 1, 10, 25, 50, 100, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1,000, 1,250, 1,500, 1,750, 2,000, 2,500, 3,000, 4,000, 5,000, or more nmole of isoprene/gram of cells for the wet weight of the cells/hour (nmole/gwcm/hr). In some embodiments, the amount of isoprene is between about 2 to about 5,000 nmole/gwcm/hr, such as between about 2 to about 100 nmole/gwcm/hr, about 100 to about 500 nmole/gwcm/hr, about 150 to about 500 nmole/gwcm/hr, about 500 to about 1,000 nmole/gwcm/hr, about 1,000 to about 2,000 nmole/gwcm/hr, or about 2,000 to about 5,000 nmole/gwcm/hr. The amount of isoprene in units of nmole/gwcm/hr can be measured as disclosed in U.S. Pat. No. 5,849,970, which is hereby incorporated by reference in its entirety, particularly with respect to the measurement of isoprene production. For example, two mL of headspace (e.g., headspace from a culture such as 2 mL of culture cultured in sealed vials at 32° C. with shaking at 200 rpm for approximately 3 hours) are analyzed for isoprene using a standard gas chromatography system, such as a system operated isothermally (85° C.) with an n-octane/porasil C column (Alltech Associates, Inc., Deerfield, Ill.) and coupled to a RGD2 mercuric oxide reduction gas detector (Trace Analytical, Menlo Park, Calif.) (see, for example, Greenberg et al, Atmos. Environ. 27A: 2689-2692, 1993; Silver et al., Plant Physiol. 97:1588-1591, 1991, which are each hereby incorporated by reference in their entireties, particularly with respect to the measurement of isoprene production). The gas chromatography area units are converted to nmol isoprene via a standard isoprene concentration calibration curve. In some embodiments, the value for the grams of cells for the wet weight of the cells is calculated by obtaining the A600 value for a sample of the cell culture, and then converting the A600 value to grams of cells based on a calibration curve of wet weights for cell cultures with a known A600 value. In some embodiments, the grams of the cells is estimated by assuming that one liter of broth (including cell medium and cells) with an A600 value of 1 has a wet cell weight of 1 gram. The value is also divided by the number of hours the culture has been incubating for, such as three hours.
In some embodiments, the cells in culture produce isoprene at greater than or about 1, 10, 25, 50, 100, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1,000, 1,250, 1,500, 1,750, 2,000, 2,500, 3,000, 4,000, 5,000, 10,000, 100,000, or more ng of isoprene/gram of cells for the wet weight of the cells/hr (ng/gwcm/h). In some embodiments, the amount of isoprene is between about 2 to about 5,000 ng/gwcm/h, such as between about 2 to about 100 ng/gwcm/h, about 100 to about 500 ng/gwcm/h, about 500 to about 1,000 ng/gwcm/h, about 1,000 to about 2,000 ng/gwcm/h, or about 2,000 to about 5,000 ng/gwcm/h. The amount of isoprene in ng/gwcm/h can be calculated by multiplying the value for isoprene production in the units of nmole/gwcm/hr discussed above by 68.1 (as described in Equation 5 below).
In some embodiments, the cells in culture produce a cumulative titer (total amount) of isoprene at greater than or about 1, 10, 25, 50, 100, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1,000, 1,250, 1,500, 1,750, 2,000, 2,500, 3,000, 4,000, 5,000, 10,000, 50,000, 100,000, or more mg of isoprene/L of broth (mg/Lbroth, wherein the volume of broth includes the volume of the cells and the cell medium). In some embodiments, the amount of isoprene is between about 2 to about 5,000 mg/Lbroth, such as between about 2 to about 100 mg/Lbroth, about 100 to about 500 mg/Lbroth, about 500 to about 1,000 mg/Lbroth, about 1,000 to about 2,000 mg/Lbroth, or about 2,000 to about 5,000 mg/Lbroth. The specific productivity of isoprene in mg of isoprene/L of headspace from shake flask or similar cultures can be measured by taking a 1 ml sample from the cell culture at an OD600 value of approximately 1.0, putting it in a 20 mL vial, incubating for 30 minutes, and then measuring the amount of isoprene in the headspace. If the OD600 value is not 1.0, then the measurement can be normalized to an OD600 value of 1.0 by dividing by the OD600 value. The value of mg isoprene/L headspace can be converted to mg/Lbroth/hr/OD600 of culture broth by multiplying by a factor of 38. The value in units of mg/Lbroth/hr/OD600 can be multiplied by the number of hours and the OD600 value to obtain the cumulative titer in units of mg of isoprene/L of broth.
The instantaneous isoprene production rate in mg/Lbroth/hr in a fermentor can be measured by taking a sample of the fermentor off-gas, analyzing it for the amount of isoprene (in units such as mg of isoprene per Lgas), and multiplying this value by the rate at which off-gas is passed though each liter of broth (e.g., at 1 vvm (volume of air/volume of broth/minute) this is 60 Lgas per hour). Thus, an off-gas level of 1 mg/Lgas corresponds to an instantaneous production rate of 60 mg/Lbroth/hr at air flow of 1 vvm. If desired, the value in the units mg/Lbroth/hr can be divided by the OD600 value to obtain the specific rate in units of mg/Lbroth/hr/OD. The average value of mg isoprene/Lgas can be converted to the total product productivity (grams of isoprene per liter of fermentation broth, mg/Lbroth) by multiplying this average off-gas isoprene concentration by the total amount of off-gas sparged per liter of fermentation broth during the fermentation. Thus, an average off-gas isoprene concentration of 0.5 mg/Lbroth/hr over 10 hours at 1 vvm corresponds to a total product concentration of 300 mg isoprene/Lbroth.
In some embodiments, the cells in culture convert greater than or about 0.0015, 0.002, 0.005, 0.01, 0.02, 0.05, 0.1, 0.12, 0.14, 0.16, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.2, 1.4, or 1.6% of the carbon in the cell culture medium into isoprene. In some embodiments, the percent conversion of carbon into isoprene is between about 0.002 to about 1.6%, such as about 0.002 to about 0.005%, about 0.005 to about 0.01%, about 0.01 to about 0.05%, about 0.05 to about 0.15%, 0.15 to about 0.2%, about 0.2 to about 0.3%, about 0.3 to about 0.5%, about 0.5 to about 0.8%, about 0.8 to about 1.0%, or about 1.0 to about 1.6%. The percent conversion of carbon into isoprene (also referred to as “% carbon yield”) can be measured by dividing the moles carbon in the isoprene produced by the moles carbon in the carbon source (such as the moles of carbon in batched and fed glucose and yeast extract). This number is multiplied by 100% to give a percentage value (as indicated in Equation 1).

% Carbon Yield=(moles carbon in isoprene produced)/(moles carbon in carbon source)*100  Equation 1

For this calculation, yeast extract can be assumed to contain 50% w/w carbon.

% Carbon Yield=(39.1 g isoprene*1/68.1 mol/g*5 C/mol)/[(181221 g glucose*1/180 mol/g*6 C/mol)+(17780 g yeast extract*0.5*1/12 mol/g)]*100=0.042%  Equation 2

One skilled in the art can readily convert the rates of isoprene production or amount of isoprene produced into any other units. Exemplary equations are listed below for interconverting between units.
Units for Rate of Isoprene Production (Total and Specific)

1 g isoprene/Lbroth/hr=14.7 mmol isoprene/Lbroth/hr(total volumetric rate)  Equation 3

1 nmol isoprene/gwcm/hr=1 nmol isoprene/Lbroth/hr/OD600(This conversion assumes that one liter of broth with an OD600 value of 1 has a wet cell weight of 1 gram.)  Equation 4

1 nmol isoprene/gwcm/hr=68.1 ng isoprene/gwcm/hr(given the molecular weight of isoprene)  Equation 5

1 nmol isoprene/Lgas O2/hr=90 nmol isoprene/Lbroth/hr(at an O2 flow rate of 90 L/hr per L of culture broth)  Equation 6

1 μg isoprene/Lgas isoprene in off-gas=60 μg isoprene/Lbroth/hr at a flow rate of 60 Lgas per Lbroth(1 vvm)  Equation 7

Units for Titer (Total and Specific)

1 nmol isoprene/mg cell protein=150 nmol isoprene/Lbroth/OD600(This conversion assumes that one liter of broth with an OD600 value of 1 has a total cell protein of approximately 150 mg) (specific productivity)  Equation 8

1 g isoprene/Lbroth=14.7 mmol isoprene/Lbroth(total titer)  Equation 9

If desired, Equation 10 can be used to convert any of the units that include the wet weight of the cells into the corresponding units that include the dry weight of the cells.

Dry weight of cells=(wet weight of cells)/3.3  Equation 10

In some embodiments encompassed by the invention, a cell comprising a heterologous nucleic acid encoding an isoprene synthase polypeptide produces an amount of isoprene that is at least or about 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, 100-fold, 150-fold, 200-fold, 400-fold, or greater than the amount of isoprene produced from a corresponding cell grown under essentially the same conditions without the heterologous nucleic acid encoding the isoprene synthase polypeptide.
In some embodiments encompassed by the invention, a cell comprising a heterologous nucleic acid encoding an isoprene synthase polypeptide and one or more heterologous nucleic acids encoding a DXS, IDI, and/or MVA pathway polypeptide produces an amount of isoprene that is at least or about 2-fold, 3-fold, 5-fold, 10-fold, 25-fold, 50-fold, 100-fold, 150-fold, 200-fold, 400-fold, or greater than the amount of isoprene produced from a corresponding cell grown under essentially the same conditions without the heterologous nucleic acids.
Exemplary Isoprene Purification Methods
In some embodiments, any of the methods described herein further include recovering the isoprene. For example, the isoprene produced using the compositions and methods of the invention can be recovered using standard techniques. such as gas stripping, fractionation, adsorption/desorption, pervaporation, thermal or vacuum desorption of isoprene from a solid phase, or extraction of isoprene immobilized or absorbed to a solid phase with a solvent (see, for example, U.S. Pat. Nos. 4,703,007 and 4,570,029, which are each hereby incorporated by reference in their entireties, particularly with respect to isoprene recovery and purification methods). In some embodiments, the recovery of isoprene involves the isolation of isoprene in a liquid form (such as a neat solution of isoprene or a solution of isoprene in a solvent). Gas stripping involves the removal of isoprene vapor from the fermentation off-gas stream in a continuous manner. Such removal can be achieved in several different ways including, but not limited to, adsorption to a solid phase, partition into a liquid phase, or direct condensation. In some embodiments, membrane enrichment of a dilute isoprene vapor stream above the dew point of the vapor resulting in the condensation of liquid isoprene.
The recovery of isoprene may involve one step or multiple steps. In some embodiments, the removal of isoprene vapor from the fermentation off-gas and the conversion of isoprene to a liquid phase are performed simultaneously. For example, isoprene can be directly condensed from the off-gas stream to form a liquid. In some embodiments, the removal of isoprene vapor from the fermentation off-gas and the conversion of isoprene to a liquid phase are performed sequentially. For example, isoprene may be adsorbed to a solid phase and then extracted from the solid phase with a solvent.
In some embodiments, any of the methods described herein further include purifying the isoprene. For example, the isoprene produced using the compositions and methods of the invention can be purified using standard techniques. Purification refers to a process through which isoprene is separated from one or more components that are present when the isoprene is produced. In some embodiments, the isoprene is obtained as a substantially pure liquid. Examples of purification methods include (i) distillation from a solution in a liquid extractant and (ii) chromatography. As used herein, “purified isoprene” means isoprene that has been separated from one or more components that are present when the isoprene is produced. In some embodiments, the isoprene is at least about 20%, by weight, free from other components that are present when the isoprene is produced. In various embodiments, the isoprene is at least or about 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 95%, or 99%, by weight, pure. Purity can be assayed by any appropriate method, e.g., by column chromatography, HPLC analysis, or GC-MS analysis.
Crystal Structure of Isoprene Synthase
The invention also contemplates crystalline forms of plant isoprene synthase (e.g., poplar and kudzu) and its variants as described supra and in the Examples. In one embodiment, the invention comprises any polypeptide which has the crystal structure of poplar isoprene synthase as disclosed in Table 16-7.
EXPERIMENTAL
The following examples are provided in order to demonstrate and further illustrate certain preferred embodiments and aspects of the present invention and are not to be construed as limiting the scope thereof.
In the experimental disclosure which follows, the following abbreviations apply: ° C. (degrees Centigrade); rpm (revolutions per minute); H2O (water); diH2O (deionized water); aa and AA (amino acid); by (base pair); kb (kilobase pair); kD (kilodaltons); gm (grams); μg and ug (micrograms); mg (milligrams); ng (nanograms); μl and ul (microliters); ml (milliliters); mm (millimeters); qs (quantity sufficient); nm (nanometers); μm and um (micrometer); M (molar); mM (millimolar); μM and uM (micromolar); pM (picomolar); U (units); MW (molecular weight); sec (seconds); min (minute/minutes); hr (hour/hours); OD600 (optical density at 600 nm); BSA (bovine serum albumin); DMAPP (dimethylallyl diphosphate); DTT (dithiothreitol); EtOH (ethanol); IPTG (isopropyl-beta-D-thiogalactopyranoside); isoprene (2-methyl-1,3-butadiene); IspS (isoprene synthase); PAGE (polyacrylamide gel electrophoresis); PBS (phosphate buffered saline [150 mM NaCl, 10 mM sodium phosphate buffer, pH 7.2]); and SDS (sodium dodecyl sulfate).
The following abbreviations apply to companies whose products or services may have been referred to in the experimental examples: Agilent (Agilent Technologies, Santa Clara, Calif.); Becton Coulter (Becton Coulter, Inc., Fullerton, Calif.); Bio-Rad (Bio-Rad Laboratories, Hercules, Calif.); Cayman Chemical (Cayman Chemical Co., Ann Arbor, Mich.); CTC Analytics (CTC Analytics A.G., Zwingen, Switzerland); EMS (Electron Microscopy Supply, Hatfield, Pa.); Epicentre (Epicentre Biotechnologies, Madison, Wis.); Integrated DNA Technologies (Integrated DNA Technologies, Coralville, Iowa); Invitrogen (Invitrogen Corp., Carlsbad, Calif.); Molecular Dynamics (Molecular Dynamics, Sunnyvale, Calif.); Novagen (Novagen, Inc., Madison, Wis.); Perkin Elmer (Perkin Elmer, Waltham, Mass.); Roche (Roche Applied Science, Indianopolis, Ind.); Sigma (Sigma-Aldrich, St. Louis, Mo.); Stratagene (Stratagene Cloning Systems, La Jolla, Calif.); Qiagen (Qiagen, Inc., Valencia, Calif.); Takara (Takara Bio USA, Madison, Wis.); Thomson Instrument (Thomson Instrument Co., Oceanside, Calif.); V&P Scientific (V&P Scientific, Inc., San Diego, Calif.); and Zinsser (Zinsser North America, Northridge, Calif.).
EXAMPLE 1
Cloning of Kudzu Isoprene Synthase for Expression in Recombinant Bacteria
In this example, methods used to produce kudzu isoprene synthase (IspS) in E. coli are described. The protein sequence for the kudzu (Pueraria montana) isoprene synthase gene (IspS) was obtained from GenBank (AAQ84170). A kudzu isoprene synthase gene, optimized for E. coli codon usage, was purchased from DNA2.0 (Menlo Park, Calif.), and is set forth as SEQ ID NO:1 (FIG. 1). The isoprene synthase gene was removed from the supplied plasmid by restriction endonuclease digestion with BspLU11I/PstI gel-purified, and ligated into pTrcHis2B (Invitrogen) that had been digested with NcoI/PstI. The construct was designed such that the stop codon in the isoprene synthase gene was 5′ to the PstI site. As a result, when the construct was expressed the His-Tag is not attached to the isoprene synthase protein. The resulting plasmid, pTrcKudzu, was verified by sequencing.
The isoprene synthase gene was also cloned into pET16b (Novagen). In this case, the isoprene synthase gene was inserted into pET16b such that the recombinant isoprene synthase protein contained the N-terminal His tag. The isoprene synthase gene was amplified from pTrcKudzu by PCR using the primer set pET-His-Kudzu-2F: 5′-CGTGAGATCA TATGTGTGCG ACCTCTTCTC AATTTAC (SEQ ID NO:3) and pET-His-Kudzu-R: 5′-CGGTCGACGG ATCCCTGCAG TTAGACATAC ATCAGCTG (SEQ ID NO:4). These primers added an NdeI site at the 5′-end and a BamH1 site at the 3′ end of the gene respectively. The plasmid pTrcKudzu, described above, was used as template DNA, HERCULASE DNA polymerase (Stratagene) was used according to manufacturer's directions, and primers were added at a concentration of 10 pM. The PCR was carried out in a total volume of 25 μl. The PCR product was digested with NdeI/BamH1 and cloned into pET16b digested with the same enzymes. The ligation mix was transformed into E. coli Top10 (Invitrogen) and the correct clone selected by sequencing. The resulting plasmid designated pETNHisKudzu is then transformed into BL21(λDE3)pLysS (Novagen) cells for expression from the T7 promoter.
The kudzu isoprene synthase gene was also cloned into the low copy number plasmid pCL1920 (Lerner and Inouye, Nucl Acids Res, 18:4631, 1990). Primers were used to amplify the kudzu isoprene synthase gene from pTrcKudzu described above. The forward primer added a HindIII site and an E. coli consensus RBS to the 5′ end. The PstI cloning site was already present in pTrcKudzu just 3′ of the stop codon so the reverse primer was constructed such that the final PCR product includes the PstI site. The sequences of the primers were: HindIII-rbs-Kudzu F: 5′-CATATGAAAG CTTGTATCGA TTAAATAAGG AGGAATAAAC C (SEQ ID NO:5) and BamH1-Kudzu R: 5′-CGGTCGACGG ATCCCTGCAG TTAGACATAC ATCAGCTG (SEQ ID NO:4). The PCR product was amplified using HERCULASE DNA polymerase (Stratagene) with primers at a concentration of 10 pM and with 1 ng of template DNA (pTrcKudzu). The amplification protocol included 30 cycles of (95° C. for 1 minute, 60° C. for 1 minute, 72° C. for 2 minutes). The product was digested with HindIII and PstI and ligated into pCL1920, which had also been digested with HindIII and PstI. The ligation mix was transformed into E. coli Top 10. Several transformants were verified by sequence analysis. The resulting plasmid was designated pCL-lac-Kudzu.
In order to remove the beta-lactamase gene, pTrcKudzu was digested with BspHI, treated with shrimp alkaline phosphatase (SAP), incubated at 65° C. for 10 min to heat kill the SAP, then end-filled by incubating with 2 units of Klenow fragment (New England BioLabs) and dNTPS. The 5 kb fragment was purified from an agarose gel and ligated to the Kan(R) gene. The Kan(R) gene was prepared by PCR amplification from pCR-Blunt-II-TOPO (Invitrogen) using primers MCM22 and MCM23 and Taq DNA polymerase according to the Manufacturer's instructions. The PCR fragment was digested with HindIII and PvuI, and end-filled using Klenow Fragment and dNTPs. The ligation mixture was transformed into E. coli Top 10 chemically competent cells and a transformant carrying a plasmid conferring kanamycin resistance, pTrcKudzu(kan), was selected on Luria Agar containing kanamycin (50 μg/ml). The sequences of the primers were: MCM22 5′-gatcaagctt AACCGGAATTGCCAGCTG (SEQ ID NO:15); and MCM23 5′-gatccgatcgTCAGAAGAACTCGTCAAGAAGGC (SEQ ID NO:16).
EXAMPLE 2
Cloning of Poplar Isoprene Synthase for Expression in Recombinant Bacteria
In this example, methods used to produce poplar isoprene synthase (IspS) in E. coli are described. The protein sequence for the poplar (Populus alba x Populus tremula) isoprene synthase (Schnitzler et al., Planta 222:777-786, 2005) was obtained from GenBank (CAC35696). A gene, codon optimized for E. coli, was purchased from DNA2.0 and is set forth as SEQ ID NO:6 (FIG. 3). The isoprene synthase gene was removed from the supplied plasmid (p9796-poplar) by restriction endonuclease digestion with BspLU11I/PstI, gel-purified, and ligated into pTrcHis2B that had been digested with NcoI/PstI. The construct is cloned such that the stop codon in the insert is before the PstI site, which results in a construct in which the His-Tag is not attached to the isoprene synthase protein. The resulting plasmid, pTrcPoplar, was verified by sequencing using commercially available primers that hybridize within the vector sequence (Forward and Reverse), as well as the primer Poplar InSeq 5′-GAGAAAATCG GTAAGGAACT GG (SEQ ID NO:8).
EXAMPLE 3
Isoprene Production in Recombinant Bacteria
In this example, methods used to produce and measure isoprene in recombinant E. coli are described.
I. Determination of Isoprene Production
For the shake flask cultures, one ml of a culture was transferred from shake flasks to 20 ml CTC headspace vials (Agilent vial Catalog No. 5188 2753, and cap Catalog No. 5188 2759). The cap was screwed on tightly and the vials incubated at the equivalent temperature with shaking at 250 rpm. After 30 minutes, the vials were removed from the incubator and analyzed as described below. In cases where isoprene production in fermentors was determined, samples were taken from the off-gas of the fermentor and analyzed directly.
The analysis was performed using an Agilent 6890 GC/MS system interfaced with a CTC Analytics CombiPAL autosampler operating in headspace mode. An Agilent HP-5MS GC/MS column (30 m×0.25 mm; 0.25 μm film thickness) was used for separation of analytes. The sampler was set up to inject 500 μL of headspace gas. The GC/MS method utilized helium as the carrier gas at a flow of 1 ml/min. The injection port was held at 250° C. with a split ratio of 50:1. The oven temperature was held at 37° C. for the 2 min duration of the analysis. The Agilent 5793N mass selective detector was run in single ion monitoring (SIM) mode on m/z 67. The detector was switched off from 1.4 to 1.7 minutes to allow the elution of permanent gases. Under these conditions, isoprene (2-methyl-1,3-butadiene) was observed to elute at 1.78 minutes. A calibration table was used to quantify the absolute amount of isoprene and was found to be linear from 1 μg/L to 200 μg/L. The limit of detection was estimated to be 50 to 100 ng/L using this method.
II. Production of Isoprene in Shake Flasks
The vectors described above were introduced into E. coli strain BL21(λDE3)pLysS (Novagen) to produce strains BL21/ptrcKudzu, BL21/pCL-lac-Kudzu and BL21/pETHisKudzu. The strains were spread for isolation onto LA (Luria agar) containing the appropriate antibiotic (50 μg/ml carbenicillin for BL21/ptrcKudzu and BL21/pETHisKudzu or 50 μg/ml spectinomycin for BL21/pCL-lac-Kudzu) and incubated overnight at 37° C. Single colonies were inoculated into 250 ml baffled shake flasks containing 20 ml Luria Bertani broth (LB) and the appropriate antibiotic. Cultures were grown overnight at 20° C. with shaking at 200 rpm. The OD600 of the overnight cultures was measured and the cultures were diluted into a 250 ml baffled shake flask containing 30 ml MAGICMEDIA expression medium (Invitrogen) containing the appropriate antibiotic to an OD600 ̃0.05. The culture was incubated at 30° C. with shaking at 200 rpm. When the OD600 ̃0.5-0.8, 400 μM IPTG was added and the cells were incubated for a further 6 hours at 30° C. with shaking at 200 rpm. At 0, 2, 4 and 6 hours after induction with IPTG, 1 ml aliquots of the cultures were collected, the OD600 was determined and the amount of isoprene produced was measured as described above.
III. Production of Isoprene from BL21/ptrcKudzu in 14 Liter Fermentation
Large-scale production of isoprene from E. coli containing the recombinant kudzu isoprene synthase gene was determined from a fed-batch culture. The recipe for the fermentation media (TM2) per liter of fermentation medium was as follows: K2HPO4 13.6 g, KH2PO4 13.6 g, MgSO4*7H2O 2 g, citric acid monohydrate 2 g, ferric ammonium citrate 0.3 g, (NH4)2SO4 3.2 g, yeast extract 5 g, 1000× Modified Trace Metal Solution 1 ml. All of the components were added together and dissolved in diH2O. The pH was adjusted to 6.8 with potassium hydroxide (KOH) and qs to volume. The final product was filter sterilized with 0.22 μM filter, but not autoclaved. The recipe for 1000× Modified Trace Metal Solution was as follows: Citric Acids*H2O 40 g, MnSO4*H2O 30 g, NaCl 10 g, FeSO4*7H2O 1 g, CoCl2*6H2O 1 g, ZnSO*7H2O 1 g, CuSO4*5H2O 100 mg, H3BO3 100 mg, NaMoO4*2H2O 100 mg. Each component was dissolved one at a time in diH2O, the pH was adjusted to 3.0 with HCl/NaOH, then qs to volume and filter sterilized with a 0.22μ filter.
This experiment was carried out in 14 L bioreactor to monitor isoprene formation from glucose at the desired fermentation, pH 6.7 and temperature 34° C. An inoculum of E. coli strain BL21/ptrcKudzu taken from a frozen vial was prepared in soytone-yeast extract-glucose medium in two 600 ml flasks. After the inoculum grew to OD550=0.6, two 600 ml flasks were centrifuged and the contents resuspended in 70 ml supernatant to transfer the cell pellet (70 ml of OD 3.1 material) to the bioreactor. At various times after inoculation, samples were removed and the amount of isoprene produced was determined as described above.
EXAMPLE 4
Selection of Sites for Improvement of Plant Isoprene Synthases
The isoprene synthases of plants were expected to be homologous to the terpene synthases. The three-dimensional structures of two homologous terpene synthases have been determined from bornyl diphosphate synthase (pdb entry 1N1B) and 5-epi-aristolochene synthase (pdb entry 5EAU). These enzymes share only 32% homology but their tertiary structure is conserved. In addition, the structures of intermediate complexes with both enzymes have shown that not only tertiary folding, but also detailed interactions in the active sites of these enzymes are highly conserved.
The kudzu and poplar isoprene synthases have higher sequence identity than was seen between the bornyl diphosphate synthase and the 5-epi-aristolochene synthase as shown in Table 4-1 below.







TABLE 4-1







Percent Identity of Various Enzymes











BDP-synthase
5EA-synthase
Kudzu IspS














Poplar IspS
40.1
32.9
54.4


Kudzu IspS
40.7
33.8


5ES synthase
31.9









A homology model of the poplar isoprene synthase has been made based on the bornyl diphosphate synthase (BDP-synthase) pdb entry 1N24 ( ̃40% sequence identity). The homology model appears to be plausible based on the close similarity of 10 trial models created using the program MOE written and supported by The Chemical Computing Group, Inc. The plausibility is based on the conservation of common amino acid residues at sites found to be involved in catalysis in the BDP-synthase structure.
A comparison of the active site from the structure of BDP-synthase and the homology model of poplar IspS indicates that the active site involved in metal ion binding and phosphate recognition is conserved. In particular, Lys 272, Asp 309, Asp 313, Glu 387, Arg 450 and Asn 453 of poplar IspS were observed to overlap equivalent residues in BDP-synthase. In this example, amino acid residue positions for poplar IspS are derived from SEQ ID NO:7. The positioning of an intermediate of the BDP-synthase was also compared with the poplar IspS homology model. Based on this, it was possible to identify the analogous binding region and the approach direction that isopentenyl diphosphate would require in order to bind and react with the poplar IspS enzyme.
A homology model of the kudzu isoprene synthase has been made based on bornyl diphosphate synthase pdb entry 1N24 having ( ̃40% sequence identity). A comparison of the active site from the structure of BDP-synthase with the homology model of kudzu IspS indicates that numerous active site residues involved in metal ion binding and phosphate recognition are conserved. In particular, Arg 269, Asp 306, Asp 310, Glu 384, Arg 450 and Asn 453 of kudzu IspS were observed to overlap equivalent residues in BDP-synthase.
A comparison of the active site residues identified in the homology models of poplar and kudzu IspS revealed that residues from one homology model are also quite homologous with similar residues, appearing with only minor shifts in the relative position numbers for some of the residues, in the other homology model. Based on the homology models, sites in poplar and kudzu IspS were identified as candidates for mutagenesis to produce variant IspS enzymes with improved performance. Briefly, sites were selected in the IspS that might alter the interaction of the metal binding, the diphosphate recognition, the IPP chain binding and/or the approach to the active site.
I. Diphosphate/Metal Binding Sites
The side chains of amino acid residues in the poplar IspS that are found in proximity to the metal and diphosphate (DPP) binding side chains were identified. These residues include Phe 384, Tyr 402, Ala 406, Ser 409, Ala 460 and Asn 469. The inventors note that Lys 272 is incorrect based on homology to other known poplar IspS sequences, which have an Arg at this position.
II. Substrate Access Loop
The substrate access loop of poplar IspS is in a region that deviates from the BDP-synthase structure. In the BDP-synthase structure the residues form a segment that creates a cover. Without being bound by theory, the inventors expect that this segment in the actual three-dimensional structure of poplar IspS will form a similar structure. As such the residues in this loop, including residues 455-466, will be in a position to alter the activity of the poplar IspS enzyme. In the poplar IspS enzyme residues 454-466 have the following sequence:









(SEQ ID NO: 9)



LASASAEIARGET.






III. Isoprenyl Binding Site

The complex of BDP-synthase and the product of the reaction, bornyl diphosphate (pdb entry 1N24), was used to identify residues in the poplar model that may modulate substrate specificity and/or reaction rate (altered on and off rates of substrate and product). These residues include Arg 274, Trp 281 Phe 302, Val 305, Ser 411, Gln 415, Phe 449, Ser 537 and Glu 540.







TABLE 4-2







Candidate Mutagenesis Sites










Poplar
Kudzu



Amino Acid
Amino Acid/codon















DPP/metal sites
Phe 384
Phe 381/1141-1143




Tyr 402
Tyr 399/1195-1197




Ala 406
Ala 403/1207-1209




Ser 409
Ser 406/1216-1218




Asn 469
Asn 469/1405-1407



Extra DPP sites
Tyr 312
Tyr 309/925-927




Asp 313
Asp 310/928-930




Leu 380
Leu 377/1129-1131




Glu 387
Glu 384/1150-1152




Asn 404
Asn 402/1204-1206




Ser 410
Ser 407/1219-1221



N_terminal access segment
22
20/58-60




23
21/61-63




24
22/64-66




25
23/67-69




26
24/70-72




27
25/73-75



Substrate access loop
Leu 454
Ala 456/4102-4104




Ala 455
Thr 457/4105-4107




Ser 456
Ser 458/4108-4110




Ala 457
Ala 459/4111-4113




Ser 458
Ala 460/4114-4116




Ala 459
Glu 461/4117-4119




Glu 460
Leu 462/4120-4122




Ile 461
Glu 463/4123-4125




Ala 462




Arg 463
Arg 464/4126-4128




Gly 464
Gly 465/4129-4131




Glu 465
Glu 466/4132-4134




Thr 466
Thr 467/4135-4137





Thr 468/4138-4140



Isoprenyl binding site
Arg 274
Arg 271/811-813




Trp 281
Trp 278/832-834




Phe 302
Phe 299/895-897




Val 305
Val 302/904-906




Ser 411
Ser 408/1222-1224




Gln 415




Phe 449
Phe 449/1345-1347




Ser 537
Ser 458/1372-1374




Glu 540
Tyr 531/1591-1593



Extra Isoprenyl sites
Gly 412
Gly 409/1225-1227




Leu 414
Ala 411/1231-1233




Leu 416
Leu 413/1237-1239




Leu 521
Met 523/1567-1569




Ser 525
Ser 527/1579-1581










EXAMPLE 5
Mutation of Non-Conserved Cysteines in Kudzu Isoprene Synthase
The kudzu and poplar isoprene synthase (IspS) homology models, based on the bornyl diphosphate synthase crystal structure, were compared with respect to the positions of the cysteine residues. Cysteines have the potential to form disulfide bonds and stabilize structures. The non-conserved cysteines, contemplated to affect solubility and/or activity, were altered by site-directed mutagenesis. The kudzu IspS amino acid sequence used for the modeling is shown in FIG. 2 (SEQ ID NO:2). There are eight cysteines in kudzu IspS at positions 57, 291, 343, 378, 421, 446, 451 and 529 (relative to the mature form of the protein) as shown in the homology model of FIG. 5. In contrast, there are five cysteines in poplar IspS amino acid sequence has five cysteines, as shown in the homology model of FIG. 6 upon which the kudzu cysteines are superimposed. Several of the cysteines are apparently conserved between the poplar and kudzu IspS sequences indicating that these positions are important in stabilizing the structure, activity and/or other protein function. The remaining cysteines (nonconserved residues 57, 291, 421 and 446) in kudzu were mutated to serine as described herein.
I. Mutagenesis
The QUIKCHANGE® Multi-Site Directed Mutagenesis Kit (Stratagene) was used as per the manufacturer's directions. The following primers were utilized for mutagenesis:









C57S-F



(SEQ ID NO: 10)



5′-CTGGAGGAAGAAGTTCGC TCC



 



ATGATCAACCGTGTAGAC;



 



C291S-F



(SEQ ID NO: 11)



5′-CGCCAGACCCGCAGTTTGGTGAA TCT



 



CGCAAAGCTGTTACTAAAATG;



 



C421S-F



(SEQ ID NO: 12)



5′-CGCCGTCTTACTTTTCCGTA TCC



 



CAGCAGCAGGAAGACATC;



 



C446S-F



(SEQ ID NO: 13)



5′-CATGGTCTGGTGCGTTCTAGC TCC



 



GTTATCTTCCGCCTGTGC;



and



 



C421S-R



(SEQ ID NO: 14)



5′-GATGTCTTCCTGCTGCTG GGA



 



TACGGAAAAGTAAGACGGCG.






The plasmid pTrcKudzu(kan) described in Example 1 was used as template DNA. The primers C57S-F, C291S-F, C421S-F, and C446-F were combined in a single reaction (100 pmol). Template DNA was added ( ̃200 nanograms) and 0.5 μl of Quiksolution was added to the recommended volumes of enzyme and buffer. The PCR reaction was carried out in an Eppendorf PCR machine using an annealing temperature of 55° C. and an extension time of 12 minutes for 30 cycles. Other parameters of the cycle were as indicated in the instructions. The PCR mix was treated with DpnI for 4 hours at 37° C. (2×1 μl for 2 h each) and then 5 μl of the reaction were transformed into E. coli Top10 (Invitrogen) chemically competent cells and plated on Luria agar containing kanamycin (50 μg/ml). After overnight incubation at 37° C., several colonies were picked and inoculated into 5 ml of Luria Broth containing kanamycin (50 μg/ml). The plasmids were isolated using the QIAprep Spin Miniprep kit (Qiagen), and the IspS genes were sequenced in their entirety. Various single and combinations of mutations were made as indicated in the Table 5-1 below.







TABLE 5-1







BL21(λDE3) Cells Transformed with Mutated pTrcKudzu Plasmids*













Strain*
C57S
C291S
C421S
C446S







C1
+
+





C2

+

+



C4
+
+

+



C6
+


+



C11



+



C20


+



C6-4
+

+
+










All the variant plasmids were transformed into chemically competent BL21(λDE3) cells (Novagen). In a second reaction pTrcKudzu(kan) and plasmid DNA isolated from C6 were used as templates in a QUIKCHANGE® site directed mutagenesis kit (Stratagene) single site reaction with C421S-F and C421S-R primers. After confirmation by sequencing, two additional strains were obtained.
II. Cell Growth and Isoprene Production
Cells were grown in 5 ml tubes containing Luria Broth supplemented with 50 mg/L kanamycin at 30° C. overnight with agitation. These cultures were diluted into TM3 broth supplemented with 10 g/L glucose and 50 mg/L kanamycin. The culture volume was 25 ml in a 250 ml baffled Bellco Delong flask in which cells were grown at 30° C. with agitation (225 rpm). Samples were taken aseptically, as indicated, for optical density measurements at A600. The results are shown in FIG. 7. The cultures were induced at 3.33 hrs with 200 μM IPTG and allowed to continue growth until harvest at 7.8 hr. The cultures were centrifuged at 10,000×g for 10 min, the supernatants decanted and the cell pellets frozen at −80° C. overnight.
Frozen cell pellets were thawed and resuspended in 2 ml PEB (50 mM Tris-HCl, pH 8.0, 20 mM MgCl, 2 mM dithiothreitol, and 50% [v/v] glycerol). Cells were lysed by French pressure cell disruption, one pass, at 20,000 psi. The lysate was centrifuged for 15 min at 10,000×g. The supernatants were decanted and the pellets resuspended in 2 ml PEB. The pellets and supernatants were analyzed by SDS-PAGE, 4-12% NuPage gels (Invitrogen), run in MES buffer under reducing conditions. The molecular weight standard was SeeBlue2 (Invitrogen). The results are shown in FIG. 8. The IspS protein concentrations were estimated using the BCA assay (Pierce) using BSA as standard (Table 5-2).
III. Assays for Isoprene Synthase Activity and Solubility
Briefly the activity of the supernatants was measured by reaction with DMAPP, and the isoprene evolved was quantified by GC/MS.
Headspace Assay. A sample of 200 μl of the desired culture is inoculated into 2 ml CTC headspace vials (Agilent vial Catalog No. 5188 2753, and cap Catalog No. 5188 2759). The cap was screwed on tightly and the vials were incubated at 37° C. with shaking at 250 rpm. After 30 minutes the vials were removed from the incubator and cooled briefly with ambient tap water. The vials were placed into the CombiPal Headspace auto sampler for analysis by GC-MS. The analysis was performed using an Agilent 6890 GC/MS system interfaced with a CTC Analytics CombiPAL autosampler operating in headspace mode. An Agilent HP-5MS GC/MS column (30 m×0.25 mm; 0.25 μm film thickness) was used for separation of analytes. The sampler was set up to inject 500 μL of headspace gas. The GC/MS method utilized helium as the carrier gas at a flow of 1 ml/min. The injection port was held at 250° C. with a split ratio of 50:1. The oven temperature was held at 37° C. for the 2 min duration of the analysis. The Agilent 5793N mass selective detector was run in single ion monitoring (SIM) mode on m/z 67. The detector was switched off from 1.4 to 1.7 minutes to allow the elution of permanent gases. Under these conditions isoprene (2-methyl-1,3-butadiene) was observed to elute at 1.78 minutes. A calibration table was used to quantify the absolute amount of isoprene and was found to be linear from 1 μg/L to 200 μg/L. The limit of detection was estimated to be 50-100 ng/L using this method.
DMAPP Assay. An aliquot of 95 μl of the supernatant fraction from the centrifuged French Pressure cell lysate was added to the headspace vials. A 5 μL aliquot of 8 mM DMAPP in 100 mM potassium phosphate buffer, pH 8.2 was added, the vials sealed and allowed to incubate at room temperature for 30 min. The amount of isoprene produced was measured by GC/MS as described above and reported in Table 5-2.







TABLE 5-2







Isoprene Synthase Activity from Crude Extracts of Cysteine Mutants













Activity
Protein
Specific Activity



Supernatant
(mU/ml)
(mg/ml)
(mU/mgP)
















Wt
59.0
2.48
23.79



C1
5.3
2.64
2.01



C2
0.2
3.52
0.06



C4
0.5
3.22
0.16



C6
9.0
3.32
2.71



C11
0.1
4.26
0.03










The values shown in Table 5-2 are averages of reactions with two different concentrations of extract. All proteins containing any of the cysteine mutations resulted in severe diminution of enzyme activity and an apparent decrease of soluble protein as judged by a relative increase in the proportion of protein in the insoluble (pellet) fraction.
EXAMPLE 6
Mutation of Residues in Poplar Isoprene Synthase
Alignment of the amino acid sequences of kudzu and poplar isoprene synthases with other synthases was done using Vector Nti (Invitrogen). The aligned sequences included: beta-ocimene synthase Lotus corniculatus (AAT86042); putative terpene synthetase Medficago trunculata (AAV36465); hypothetical protein Vitis vinifera (can65805); hypothetical protein Vitis vinifera (CAN62729); pinene synthase Quercus ilex (CAK55186); IspS Pueraria montana (kudzu) Sharkey (AAQ84170); monoterpene synthase Eucaliptus globulus (BAF02832); IspS Populus nigra Fortunati (CAL69918); IspS Populus tremuloides Sharkey (AAQ16588); IspS Populus alba (BAD98243); and IspS Populus alba x tremula Zimmer (CAC35696). The sequence from the database of Populus alba x tremula (CAC35696) exhibited different amino acids at positions 272 and 497 that were otherwise highly conserved. Additionally based on analysis of the homology model of poplar IspS, position 453 was identified as a third candidate for mutagenesis.
I. Mutagenesis
The QUIKCHANGE® Multi-Site Directed Mutagenesis kit (Stratagene) was used as per the manufacturer's directions to introduce the following mutations singly and in combination into the Populus alba x tremula IspS sequence (SEQ ID NO:7): K272R; C497W; and N453D. The following primers were utilized for mutagenesis:







Poplar K272R


(SEQ ID NO: 17)


5′-ccaaactgcacttcg ctcgtgaccgcctgattgag;


 


Poplar N453D


(SEQ ID NO: 18)


5′-atctttcgcctgtgcgacgacctggcaagc;


and


 


Poplar C497W


(SEQ ID NO: 19)


5′-tgaatctgatcgacgaaacctggaagaaaatgaacaaagaaaaac.






The following primer, Poplar InSeq, 5′-gagaaaatcggtaaggaactgg (SEQ ID NO:8) was used for sequencing.

Mutagenesis was done according to the manufacturer's directions, with all three mutagenesis primers being added to a single reaction mix (100 ng each) with pTrcPoplar as the template DNA (100 ng). Addition of 0.5 μl of Quik Solution aided the mutagenesis reaction. The suggested PCR cycle was run with an annealing temperature of 55° C. and an extension time of 12 min. Other parameters were as indicated in the instructions. The PCR mix was digested with DpnI for 4 hrs at 37° C. (1 μl each×2 h) and then 5 μl of the reaction was transformed into E. coli Top 10 chemically competent cells (Invitrogen). Several colonies were selected and grown up in 5 ml of Luria Broth (LB) containing carbenicillin (50 μg/ml). Plasmids were isolated using the Qiagen QIAprep spin mini-prep kit and sent for sequencing using forward and reverse primers that hybridized to the vector, as well as the Poplar InSeq primer.
The resulting variants of pTrcPoplar were obtained: pTrcPoplar K272R, pTrcPoplar K272R/N453D; pTrcPoplar K272R/N453D/C497W; and pTrcPoplar 272R/497W. These plasmids were transformed into BL21(λDE3)pLysS chemically competent cells (Novagen) for analysis. The variants were analyzed for headspace activity (production of isoprene from whole cells), solubility, and specific activity.
II. Cell Growth and Isoprene Production
The variants, the parent Poplar strain, and the strain containing pTrcKudzu were grown overnight at 37° C. in 5 ml of Luria Bertani medium containing either carbenicillin (50 μg/ml-Poplar strains) or kanamycin (50 μg/ml-Kudzu strain). These cultures were diluted into TM3 broth to an OD600 of 0.05, supplemented with 10 g/L glucose and either 50 μg/ml carbenicillin (Poplar mutants and wild type) or 50 mg/L kanamycin (BL21/pTrcKudzu). The recipe for TM3 broth is as follows: K2HPO4 (13.6 g/l) KH2PO4 (13.6 g/l), MgSO4*7H2O (2 g/l) Citric Acid Monohydrate (2 g/L) Ferric Ammonium Citrate (0.3 g/L) (NH4)2SO4 (3.2 g/L) yeast extract (0.2 g/L) 1 ml of 1000× Trace Elements solution, pH adjusted to 6.8 with ammonium hydroxide qs to volume with sterile diH2O and filter sterilized with a 0.22 micron filter. The recipe for 1000× Trace Elements solution is as follows: Citric Acids*H2O (40 g/L), MnSO4*H2O (30 g/L), NaCl (10 g/L), FeSO4*7 H2O (1 g/L), CoCl2*6 H2O (1 g/L), ZnSO*7 H2O (1 g/L), CuSO4*5 H2O (100 mg/L), H3BO3 (100 mg/L), NaMoO4*2 H2O (100 mg/L). Each component was dissolved one at a time in diH2O, pH adjusted to 3.0 with HCl/NaOH, qs to volume and filter sterilized with a 0.22 micron filter.
The diluted culture volume was 25 ml in a 250 ml baffled Bellco Delong flask for growth at 30° C. with agitation (225 rpm). Samples were taken aseptically, as indicated, for optical density measurements at A600. Two sets of cultures were set up, one for induction with 0.2 mM IPTG and one that remained un-induced. After 3 hours of growth at 30° C. with shaking at 200 rpm (OD600 ̃0.5), one set of the cultures was induced with 0.2 mM IPTG and incubated for a further 3 h at 30° C. with shaking at 200 rpm, the un-induced set was further incubated for the same amount of time. The OD600 was determined for all cultures prior to the induction time (3 h post inoculation) and at the time of the measurement of isoprene by the Headspace assay (3 h post-induction, total of 6 h of growth). The cell cultures were centrifuged at 7000×g for 15 minutes in a Sorvall superspeed centrifuge to pellet the cells. The supernatant was removed and the cell pellet frozen for use in an in vitro assay for isoprene synthase activity. Results of the growth and headspace assays are shown in the following tables.







TABLE 6-1







Growth of E. coli Strains Expressing Poplar IspS Variants


(OD600 values)











Pre-induction
Post-induction
Non-induced



(after 3 h
(3 h with
(after 6 h


Strain
growth)
IPTG)
growth)













pTrcPoplar
0.49
3.3
4.7


K272R


pTrcPoplar
0.48
3.7
4.8


K272R/N453D


pTrcPoplar
0.41
3.1
5.1


K272R/N453D/C497W


pTrcPoplar
0.45
0.8
0.97


K272R/497W H2


pTrcPoplar
0.49
3.5
4.7


WT


pTrcKudzu
0.44
3.3
4.2


WT
















TABLE 6-2







Production of Isoprene by E. coli Strains Expressing


Poplar IspS Variants (μg/L)











Pre-induction
Post-induction
Non-induced



(after 3 h
(3 h with
(after 6 h


Strain
growth)
IPTG)
growth)













pTrcPoplar
n/a
0
0


K272R


pTrcPoplar
n/a
0
0


K272R/N453D


pTrcPoplar
n/a
0
0


K272R/N453D/C497W


pTrcPoplar
n/a
0
0


K272R/497W H2


pTrcPoplar
n/a
0
0


WT


pTrcKudzu
n/a
1.3
0.29


WT









Surprisingly, the strain expressing the kudzu IspS demonstrated any measurable isoprene production. This is unexpected given that kinetic properties of the poplar enzymes were reported in the literature to be superior to those of kudzu enzymes. In particular, the prior art describes the specific activity (U/mg) and Km (μM) of recombinant kudzu IspS to be 0.075 and 7,700 respectively, native aspen IspS to be 0.5 and 8,000 respectively, and recombinant poplar IspS to be 0.16 and 9,000 respectively (Silver and Fall, J Biol Chem, 270:13010-1316, 1995; Miller et al., Planta, 213:483-487, 2001; and Sharkey et al., Plant Physiology, 137:700-712, 2005). The published Km values for the three enzymes are all quite high and within range of each other, but the specific activity for kudzu isoprene synthase is significantly worse than that of the other two isoprene synthases.
III. Assays for Isoprene Synthase Activity and Solubility
By using the DMAPP assay, the activity of isoprene synthase can be measured directly as DMAPP is the direct substrate for the enzyme. The cell pellets of the poplar parent and mutant strains, as well as the wild type kudzu were thawed and resuspended in 2 ml PEB (50 mM Tris-HCl, pH 8.0, 20 mM MgCl, 2 mM dithiothreitol, and 50% [v/v] glycerol). Cells were lysed by French pressure cell disruption, one pass, at 20,000 psi. The lysate (1 ml) was then centrifuged in a microfuge for 20 min at 20,000 rpm at 4° C. The supernatant was removed and the pellet resuspended in 1 ml of PEB. The supernatant and pellet samples were analyzed by SDS-PAGE, and DMAPP assay, while the total protein content was determined by BCA.







TABLE 6-3







DMAPP Assay of Isoprene Production from the


Supernatant of the Centrifuged Cell Lysate











OD600
Total Protein
Isoprene/


Strain
(prior to lysis)
(mg/ml)
Total Protein













Induced





pTrcPoplar
3.3
1.70
0.03


K272R


pTrcPoplar
3.6
1.3
0.08


K272R/N453D


pTrcPoplar
3.1
1.10
0.08


K272R/N453D/C497W


pTrcPoplar
0.803
0.80
0.11


K272R/497W H2


pTrcPoplar
3.5
1.30
0.06


WT


pTrcKudzu
3.3
1.50
11.15


WT


Uninduced


pTrcPoplar
4.7
1.7
0.26


K272R


pTrcPoplar
4.8
1.8
0.07


K272R/N453D


pTrcPoplar
5.2
1.9
0.02


K272R/N453D/C497W


pTrcPoplar
0.969
1.0
0.17


K272R/497W H2


pTrcPoplar
4.6
1.7
0.05


WT


pTrcKudzu
4.2
1.9
1.62


WT









Production was normalized to total cell lysate supernatant protein.







TABLE 6-4







DMAPP Assay of Isoprene Production from the


Pellet of the Centrifuged Cell Lysate











OD600
Total Protein
Isoprene/


Strain
(prior to lysis)
(mg/ml)
Total Protein













Induced





pTrcPoplar
3.3
1.02
0.118


K272R


pTrcPoplar
3.6
1.36
0.000


K272R/N453D


pTrcPoplar
3.1
1.49
0.040


K272R/N453D/C497W


pTrcPoplar
0.80
1.38
0.043


K272R/C497W H2


pTrcPoplar
3.5
1.57
0.050


WT


pTrcKudzu
3.3
1.47
0.040


WT


Uninduced


pTrcPoplar
4.7
1.40
0.170


K272R


pTrcPoplar
4.8
1.53
0.120


K272R/N453D


pTrcPoplar
5.2
1.42
0.131


K272R/N453D/C497W


pTrcPoplar
0.97
1.55
0.080


K272R/C497W H2


pTrcPoplar
4.6
1.38
0.120


WT


pTrcKudzu
4.2
1.55
0.120


WT









Production was normalized to total cell lysate pellet protein.
The poplar variant K272R/C497W showed a 1.8× increase in activity as compared to the wild type in the supernatant fraction of the induced cultures. Likewise, the poplar variants K272R and K272R/C497W showed a 5.2× and 3.4× increase in activity as compared to the wild type in the supernatant fraction of the uninduced cultures. Moreover the poplar variant K272R showed a 2× increase in activity as compared to wild type in the pellet of the induced cultures. However, the most striking result was that the kudzu IspS is more active than the poplar IspS employed herein (185×). In the above tables, H2 is the name of the clone designated pTrcPoplar K272R/C497W H2.
EXAMPLE 7
Subcloning of Kudzu Isoprene Synthase
In this Example, methods used in the construction of kudzu isoprene synthase (IspS) SELs are described. To create an expression vector for construction of site evaluation libraries (SEL), the kudzu isoprene synthase gene was subcloned into the pET24d vector (Novagen) from the pCR2.1 vector (Invitrogen). The kudzu IspS gene was amplified from pTrcKudzu template DNA using primers MCM50 5′-GATCATGCAT TCGCCCTTAG GAGGTAAAAA AACATGTGTG CGACCTCTTC TCAATTTACT (SEQ ID NO:20); and MCM53 5′-CGGTCGACGG ATCCCTGCAG TTAGACATAC ATCAGCTG (SEQ ID NO:21). PCR reactions were carried out using Taq DNA Polymerase (Invitrogen), and the resulting PCR product was cloned into pCR2.1-TOPO TA cloning vector (Invitrogen), and transformed into E. coli Top10 chemically competent cells (Invitrogen). Transformants were plated on L agar containing carbenicillin (50 μg/ml) and incubated overnight at 37° C. Five ml Luria Broth cultures containing carbenicillin 50 μg/ml were inoculated with single transformants and grown overnight at 37° C. Five colonies were screened for the correct insert by sequencing of plasmid DNA isolated from 1 ml of liquid culture (Luria Broth) and purified using the QIAprep Spin Mini-prep Kit (Qiagen). The resulting plasmid, designated MCM93, contains the kudzu IspS coding sequence in a pCR2.1 backbone (FIG. 9). The sequence of MCM93 (SEQ ID NO:22) is shown in FIG. 10.
The kudzu coding sequence was removed by restriction endonuclease digestion with PciI and BamH1 (Roche) and gel purified using the QIAquick Gel Extraction kit (Qiagen). The pET24d vector DNA was digested with NcoI and BamHI (Roche), treated with shrimp alkaline phosphatase (Roche), and purified using the QIAprep Spin Mini-prep Kit (Qiagen). The kudzu IspS fragment was ligated to the NcoI/BamH1 digested pET24d using the Rapid DNA Ligation Kit (Roche) at a 5:1 fragment to vector ratio in a total volume of 20 μl. A portion of the ligation mixture (5 μl) was transformed into E. coli Top 10 chemically competent cells and plated on L agar containing kanamycin (50 μg/ml). The correct transformant was confirmed by sequencing and transformed into chemically competent BL21(λDE3)pLysS cells (Novagen). A single colony was selected after overnight growth at 37° C. on L agar containing kanamycin (50 μg/ml). A map of the resulting plasmid designated as pET24D-Kudzu is shown in FIG. 11. The sequence of pET24D-Kudzu (SEQ ID NO:23) is shown in FIG. 12. IspS Activity was confirmed using the headspace assay as described in Example 5).
EXAMPLE 8
Construction of Isoprene Synthase Site Evaluation Libraries (SELs)
In order to improve the kinetic parameters of a plant IspS SELs are prepared at sites selected from homology models of both the kudzu and the poplar IspS enzymes. While it is predicted from the homology models that engineering at the indicated sites would improve both enzymes, in this embodiment, kudzu SELs are described. Kudzu IspS surface sites of interest include but are not limited to: 26 L, 30 E, 31 F, 33 Q, 35 L, 36 E, 37 N, 39 L, 40 K, 41 V, 43 K, 44 L, 61 R, 62 V, 63 D, 65 Q, 87 K, 94 E, 95 N, 99 L, 100 D, 105 N, 137 K, 138 E, 143 G, 144 E, 182 N, 184 L, 185 K, 187 G, 189 N, 190 T, 225 P, 226 H, 247 K, 257 T, 258 E, 259 M, 266 D, 334 N, 353 D, 357 S, 358 I, 361 E, 389 N, 392 I, 393 I, 398 K, 401 E, 421 C, 423 Q, 424 Q, 425 E, 426 D, 430 H, 432 L, 433 R, 434 S, 437 D, 443 R, 462 L, 463 E, 476 H, 478 N, 479 D, 485 Q, 508 D, 513 P, 515 A, 532 Q, 533 Y, 537 L, 538 G, 539 R, 542 Y, 543 A, and 557 P. Kudzu IspS active site positions of interest include but are not limited to: 24 P, 25 N, 309 Y, 310 D, 377 L, 381 F, 384 E, 399 Y, 402 N, 403 A, 406 S, 407 S, 409 G, 411 A, 413 L, 449 F, 456 A, 457 T, 458 S, 458 S, 459 A, 460 A, 461 E, 462 L, 463 E, 464 R, 465 G, 466 E, 467 T, 468 T, 469 N, 523 M, 527 S, and 531 Y. Additional kudzu IspS active site positions of interest include but are not limited to: 20 A, 21 N, 22 Y, 23 Q, 271 R, 278 W, 299 F, 302 V, and 408 S. Each library SEL contains clones, maximally including 20 different variants. For example, kudzu isoprene synthase SEL 531 contains variants in which the DNA triplet coding for tyrosine at position 531 of the mature kudzu enzyme is replaced by another DNA triplet encoding: alanine, aspartic acid, cysteine, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, valine, or tryptophan. Briefly, DNA triplets of specific positions in the DNA coding strand of the mature IspS are replaced. The mutated IspS nucleic acids are subsequently ligated to a suitable expression vector and used to transform suitable host cells.
Site evaluation libraries are created either by ordering synthetic constructs (e.g., DNA2.0) or by ordering primers with the “nns” sequence in place of the codon to be mutated. The primers are then be used to mutate the gene to produce an SEL at the indicated site using commercially available mutagenesis kits (e.g., Stratagene) as has been described (e.g., WO0507682A2). The mutated codons are identified by sequence analysis. The site libraries are arrayed in 96 well master plates, and frozen for later use. Cultures are grown from the master plates and prepared for screening.
The desired end products are IspS enzymes that function optimally in a host metabolically engineered to maximize carbon flow through IspS. To this end several stages of screening are used to ensure that correct parameters are being addressed. Exemplary screens include but are not limited to: expression, DMAPP feeding for production of HG, microreactor, protein determination, and headspace assays. Expression screen: One example of a method to analyze the level of protein expression is as follows. Soluble and insoluble fractions of cell lysates (obtained from lysed cell cultures) are prepared by centrifugation. The resulting supernatants and pellets are analyzed by SDS-PAGE. The percent soluble protein is determined by densitometry analysis of the protein present in the supernatant versus the pellet.
In an exemplary embodiment, kudzu site evaluation libraries are constructed in the pET24D expression vector. The pET24D-Kudzu vector, containing the kudzu isoprene synthase gene, serves as the template DNA.
Materials:


    pET24D-Kudzu vector ( ̃50 ng/μl)
    Kudzu IS site-directed mutagenic primers (Integrated DNA Technologies)
    QUIKCHANGE® Multi Site-Directed Mutagenesis Kit (Stratagene)
    MJ Research PTC-200 Peltier Thermal Cylcer (Bio-Rad Laboratories)
    One Shot TOP10 competent cells (Invitrogen)
    QIAprep Spin Miniprep Kit (Qiagen)
    BL21(λDE3) pLysS competent cells (Invitrogen)
    Luria Broth (LB) agar plates

Methods:



The method of mutagenesis was based on the codon-specific mutation approach, in which the creation of all possible mutations in a specific DNA triplet was performed using a single forward primer with a length of 25 to 45 nucleotides, enclosing a specific designed triple DNA sequence NNS(N=A, C, T or G and S=C or G) corresponding with the sequence of the codon to be mutated. This method results in the random incorporation of nucleotides at a specific pET24D-kudzu codon of interest. Table 8-1 lists the oligonucleotide primers used for mutagenesis, with the number in the primer name corresponding with the codon position in the mature kudzu isoprene synthase enzyme sequence. All oligonucleotide primers were synthesized (Integrated DNA Technologies) on a 100 nmole scale and PAGE purified.







TABLE 8-1 







Kudzu IspS Codon-Specific Mutation Primers









Name
SEQ ID
Primer Sequence





IS_A20
NO: 24
CATAATTCCCGTCGTTCCNNSAACTATCAGC




CAAACCTG


 


IS_N21
NO: 25
CATAATTCCCGTCGTTCCGCANNSTATCAGC




CAAACCTGTG


 


IS_Y22
NO: 26
CCCGTCGTTCCGCAAACNNSCAGCCAAACCT




GTGGAATTTC


 


IS_Q23
NO: 27
GTCGTTCCGCAAACTATNNSCCAAACCTGTG




GAATTTC


 


IS_R271
NO: 28
CTGGATTTTGTACGCGACNNSCTGATGGAAG




TTTATTTC


 


IS_W278
NO: 29
CTGATGGAAGTTTATTTCNNSGCACTGGGTA




TGGCGCC


 


IS_F299
NO: 30
CAAAGCTGTTACTAAAATGNNSGGTCTGGTG




ACGATCATC


 


IS_V302
NO: 31
CTAAAATGTTTGGTCTGNNSACGATCATCGA




TGACGTG


 


IS_S408
NO: 32
GAAAACGCCAGCGTTTCCTCCNNSGGTGTAG




CGCTGCTGGC









A PCR reaction was set up in a 0.5 ml thin-walled PCR tube following the manufacturer's protocol for the QUIKCHANGE® Multi Site-Directed Mutagenesis Kit (Stratagene): 1 μl pET24 Kudzu vector (50 ng/μl); 1 μl Kudzu IS site-directed forward mutagenic primer (10 μM); 2.5 μl 10× QUIKCHANGE® Multi Reaction buffer; 1 μl dNTP Mix, 1 μl QUIKCHANGE® Multi enzyme blend (2.5 U/μl); and 18.5 μl distilled autoclaved water to provide a 25 μl total reaction mix. The pET24 Kudzu SELs were amplified using the following conditions: 95° C., for 1 min (1st cycle only), followed by 95° C. for 1 min, 55° C. for 1 min, 65° C. for 12 min, and repeat cycling 29 times. Then the reaction mixture was subjected to DpnI digestion (supplied with QUIKCHANGE® Multi Site-Directed Mutagenesis Kit) by addition of 1.5 μl DpnI restriction enzyme to each tube, and incubated at 37° C. for 2 hours to digest the parental pET24D-kudzu vector. The DpnI-treated PCR reaction was then transformed into One Shot TOP10 competent cells (Invitrogen), plated onto LB agar plates containing 50 μg/ml kanamycin, and incubated overnight at 37° C. The next day, 96 random colonies were picked and sequenced to identify a minimum of 15 of the possible 19 amino acid variants. Upon identification of the site-directed variants, each variant clone was then inoculated in a 5 ml tube of LB+50 μg/ml kanamycin and grown overnight at 37° C. with shaking (250 rpm). The following day plasmid DNA was purified using the QIAprep Spin Miniprep Kit (Qiagen). The variants were then transformed into One Shot BL21(λDE3) pLysS competent cells (Invitrogen) for protein expression screening, plated on LB agar plates containing 50 μg/ml kanamycin and 30 μg/ml chloramphenicol and incubated overnight at 37° C.
An alternative method for producing pET24D-Kudzu SELs in E. coli BL21(λDE3) pLysS cells was also successfully employed. The TOP10 competent cell transformants obtained from the DpnI-treated PCR reaction described above were harvested by applying 3 ml of LB media to the top of the agar and resuspending the cells by scraping with a sterile plate spreader. The 3 ml of pooled, resuspended cells were then used to inoculate a 25 ml shake flask containing LB+50 μg/ml kanamycin. The pooled culture was then grown overnight at 37° C. with shaking (250 rpm). The following day plasmid DNA was purified from the pooled cultures using the QIAprep Spin Miniprep Kit (Qiagen). The pooled plasmid DNA was then transformed into One Shot BL21(λDE3) pLysS competent cells for protein expression screening as described above.
To make a master plate, the correct constructs are arrayed in quadruplicate in 96 well plates. One colony of the correct sequence is used to inoculate 4 wells and the plates are grown for several hours to overnight at 37° C. in LB containing 50 μg/ml kanamycin with shaking (200 rpm). Sterile glycerol is added to the cultures to a final concentration of 15% (for a final total volume of 150-200 μl/well). The plates are then sealed using BREATHE-EASIER (EMS Catalog No. 70536-20) membranes and stored at −80° C.
EXAMPLE 9
Production and Purification of Isoprene Synthase Inclusion Bodies
Inclusion bodies containing kudzu isoprene synthase were formed when the enzyme is overexpressed in the presence of the chaperone GroELS in the strain BL21(λDE3). Briefly pETNHisKudzu (U.S. Application No. 61/013,574, herein incorporated by reference) was subcloned into pGro7 (Takara Catalog No. 3340) according to the manufacturer's instructions. A 500 mL of culture was grown essentially as described (Whittington et al., Proc Natl Acad Sci USA, 99:15375-15380, 2002). Despite the presence of chaperone and low temperature of cultivation the culture yielded predominantly inclusion bodies and only low levels of soluble active protein. The inclusion bodies were harvested using the IFOLD Protein Refolding System (Novagen Catalog No. 71552-3) according to the manufacturer's instructions. This procedure led to a high yield (>50 mg) of recombinant kudzu isoprene synthase. The purity of the inclusion body is shown in FIG. 13. This preparation was used for the production of rabbit polyclonal anti-isoprene synthase antisera (Invitrogen).
EXAMPLE 10
High Throughput Biochemical Screen of Isoprene Synthase Variants
This example describes high throughput methods for the determination of isoprene synthase activity. Libraries of BL21(λDE3)pLysS E. coli host cells capable of expressing isoprene synthase variants are arrayed in 96-well plates and stored frozen at −80° C. as 15% glycerol stocks as described above in Example 8. To analyze a plate of up to 96 variants, a replica stamp of the glycerol stock master plate is made with a 96-pin MULTI-BLOT floating pin tool (V&P Scientific Catalog No. VP 408AF) onto Luria broth agar containing appropriate antibiotic(s) (e.g., 30 μg/mL chloramphenicol, 50 μg/mL kanamycin). The replica plate is incubated over night at 30° C. to allow growth of bacterial patches. Using the same floating pin replicator a 96-square deep well plate containing 250 μL of TM3 medium supplemented with 0.08% Biospringer yeast extract and 1% glucose plus antibiotics (30 μg/mL chloramphenicol, 50 μg/mL kanamycin) is inoculated from the agar plate and incubated overnight at 30° C. The recipe for TM3 broth is as follows: K2HPO4 (13.6 g/l) KH2PO4 (13.6 g/l), MgSO4*7H2O (2 g/l) Citric Acid Monohydrate (2 g/L) Ferric Ammonium Citrate (0.3 g/L) (NH4)2SO4 (3.2 g/L) yeast extract (0.2 g/L) 1 ml of 1000× Trace Elements solution, pH adjusted to 6.8 with ammonium hydroxide qs to volume with sterile diH2O and filter sterilized with a 0.22 micron filter. The recipe for 1000× Trace Elements solution is as follows: Citric Acids*H2O (40 g/L), MnSO4*H2O (30 g/L), NaCl (10 g/L), FeSO4*7 H2O (1 g/L), CoCl2*6 H2O (1 g/L), ZnSO*7 H2O (1 g/L), CuSO4*5 H2O (100 mg/L), H3BO3 (100 mg/L), NaMoO4*2 H2O (100 mg/L). Each component was dissolved one at a time in diH2O, pH adjusted to 3.0 with HCl/NaOH, qs to volume and filter sterilized with a 0.22 micron filter. The overnight cultures are diluted with the same medium to an OD600 of 0.05 and grown in another 96-square deep well plate (Thomson Instrument, Catalog No. 951652C), with each well containing 600 μL of the dilution. The dilutions are grown at 30° C. with shaking to an OD600 of 0.8 and are then induced with IPTG added to a concentration of 400 μM. The plate is grown for 5 hours and OD600 is determined for quality control and normalization.
A volume of 400 μL of culture is transferred into a new 96-well plate (Perkin Elmer, Catalog No. 6008290) and cells are harvested by centrifugation in a Beckman Coulter Allegra 6R centrifuge at 2500×g. The pellet is resuspended in 200 μL of hypotonic buffer (5 mM MgCL2, 5 mM Tris HCl, 5 mM DTT pH 8.0) and the plate is frozen at −80° C. for a minimum time of 60 min. Cell lysate is prepared by thawing the plate and adding 32 μL of isoprene synthase DMAPP assay buffer (57 mM Tris HCl, 19 mM MgCl2, 74 μg/mL DNase I (Sigma Catalog No. DN-25), 2.63×105 U/mL of READYLYSE lysozyme solution (Epicentre Catalog No. R1802M), and 5 mg/mL of molecular biology grade BSA. The plate is incubated with shaking at 25° C. for 30 min and then placed on ice. For isoprene production, an 80 μL aliquot of lysate is transferred to a 96-deep well glass plate (Zinsser Catalog No. 3600600) and 20 μL of a 10 mM DMAPP solution in 100 mM KHPO4, pH 8.2 (Cayman Chemical Catalog No. 63180) is added. The plate is sealed with an aluminum plate seal (Beckman Coultor Catalog No. 538619) and incubated with shaking at 30° C. of 60 minutes. The enzymatic reactions are terminated by heating the glass block (70° C. for 5 min). The headspace of each well is quantitatively analyzed as described in Example 5.
To determine protein concentration 5 μL or more of lysate is run on precast gels (Invitrogen Catalog No. NP0301BOX) for western blot analysis after transfer to a nitrocellulose membrane (Invitrogen Catalog No. LC2000). The primary antibody employed is an anti-isoprene synthase antibody of Example 9. Primary antibody binding is followed by development with a secondary antibody labeled with Alexa Fluor 488 (Invitrogen Catalog No. A-11008) to permit quantitative signal determination. The western blot procedure was carried out as described by Invitrogen. The fluorescence signal was recorded with a Molecular Dynamics STORM instrument using the blue filter setting and quantitatively analyzed with the Molecular Dynamics IMAGEQUANT image analysis software package. Specific activity of the library members was calculated from the ratio of the amount of isoprene produced divided by either the A600 of the induction cultures or the isoprene synthase protein concentration determined by western blot. Isoprene synthase protein standard was calibrated by standard gel densitometry with BSA stained with Coomassie brilliant blue R250 serving as primary standard. Increased, decreased, or no change in specific activity of the entire library was tabulated for further analysis. FIG. 14 provides graphs showing isoprene synthase activity of kudzu site evaluation library (SEL) members for positions Y22, A20, and S408. Most members show highly decreased activity relative to wild type, while conservative substitutions show a lesser decrease in activity. Activity of variant A20G approximates that of the wild type kudzu enzyme, indicating that it is a candidate partner for a combinatorial mutant. Interestingly, variant S408D of library S408 showed an increase in activity compared to wild type thus providing another candidate partner for a combinatorial mutant.
EXAMPLE 11
Isoprene Synthase Truncations
This example describes the identification of the amino acid sequence of the protein in the lower band of the doublet seen in purified poplar IspS preparations (see FIG. 21). A series of N-terminally truncated IspS molecules based on putative cleavage sites identified by mass spectrometry was also generated. A shorter N-terminal truncation of IspS (the “MEA” truncation in pDu39, see below) was also generated, to examine the effect of further truncation on IspS activity (Williams D C, McGarvey D J, Katahira E J, Croteau R (1998) Biochemistry 37:12213-12220).
I. Construction of an N-Terminally 6×His-tagged IspS (in pDu27) for protein purification:
The full length P. alba IspS from the template P. alba pET24a (FIGS. 19 and 20) was prepared by PCR. The following PCR reaction was prepared: 1 μl (Template)-P. alba pET24a, 5 μl 10× PfuUltraII Fusion buffer, 1 μl dNTP's (10 mM), 1 μl primer (50 μM) primer F-(MCM219), 1 μl primer (50 μM) primer R-(MCM182), 41 μl diH2O and 1 μl of PfuUltra II Fusion DNA Polymerase (Stratagene). PCR cycling parameters were as follows: 95° C. 1 min., 95° C. 1 min, 55° C. 20 sec., 72° C. 27 sec. for 29 cycles followed by 72° C. 3 min and 4° C. until cool, using an Eppendorf Mastercycler. The PCR product was gel extracted and purified, using 0.8% E-gel (Invitrogen) and Qiagen QIAquick Gel Extraction and QIAprep Spin Miniprep kits, according to the manufacturer's recommended protocol. A 3 μl aliquot of purified product was ligated to the pET200D/TOPO vector (Invitrogen), according to the manufacturer's protocol. The reaction was incubated for 5 minutes at room temperature, and the 6 μl topoisomerase mixture was then transformed into E. coli Top10 chemically competent cells (Invitrogen) according to the manufacturer's protocol. Transformants were selected for on LB plates containing kanamycin (50 μg/ml) (Kan50), and incubated at 37° C. overnight. Five colonies were picked and screened using PuReTaq Ready-To-Go PCR Beads (Amersham) using the T7 Forward and MCM182 primers. Clones harboring inserts of the correct size were further verified by sequencing using the T7 Forward and T7 Reverse primers (Quintara Biosciences). One construct, pDu27 (FIGS. 16-18), was chosen for further study. A 1 μl aliquot of the plasmid preparation was transformed into BL21(λDE3)pLysS (Invitrogen) according to the manufacturer's protocol. Transformants were selected for on LB plates containing Kan50+ and chloramphenicol (35 μg/ml) (Cm35) and incubated at 37° C. overnight. The resulting strain was used for expression and purification of N-terminally 6×His-tagged P. alba IspS.
II. Purification of 6×His-tagged IspS
Expression of 6×His-tagged IspS
N-terminally 6×His-tagged IspS was expressed and purified from strain MD08-99. The growth procedure is suitable for histidine tagged enzymes expressed in BL21(λDE3)pLysS cells. A 10 ml of overnight culture was prepared for each 1L of planned growth. The appropriate antibiotics (50 mg/ml kanamycin, 50 mg/ml chloramphenicol, and/or 50 mg/ml Carbenecillin) was added to 10 ml of LB medium in a 25 ml flask and was inoculated with 1 colony from a fresh plate of cells or directly from glycerol frozen cell stock. Cultures were grown at 30° C. overnight with shaking at  ̃220 rpm. Day cultures were prepared in 1 liter of LB medium with appropriate antibiotics for each culture. Each 1L day culture was inoculated with 10 ml of overnight culture and grown at 30-37° C. with shaking at  ̃220 rpm until the OD600 reached  ̃0.4-0.6. Day cultures were then induced with 400 μM IPTG and allowed to continue growing at 30° C. with shaking at 220 rpm for  ̃5-6 hours. Cells were then harvested by centrifugation at 10,000×g for 10 min, 4° C. Following Harvest, cells were used directly or stored at −80° C. until ready to process.
Purification of 6×His-Tagged IspS
For purification of histidine tagged enzymes from BL21(λDE3)pLysS cells, cells were gently resuspended in fresh Lysis buffer (Lysis buffer: Ni wash buffer+0.5 mM PMSF, 0.01% Tween-20, 1 mg/ml lysozyme, 0.2 mg/ml DNaseI; Ni wash buffer: 50 mM NaH2PO4, 300 mM NaCl, 20 mM Imidazole, pH 8.0). Approximately 40-50 ml of lysis buffer was used per 1L of cell pellet. Cells were then incubated on ice for approximately 30 min. The cell suspension was then lysed fully by passing 2-3 times through a french pressure cell (large french press cell at 1200 psi/High setting) until lysate started to look clear. A sample of the lysate was saved for activity assay and gel analysis ( ̃100 μl). The lysate was then clarified by centrifuging the lysate at 30,000×g for 30 min, 4° C. in a Sorvall Discovery 90SE ultracentrifuge. The supernatant was removed and retained. A sample of the “clarified lysate” was saved for activity assay and gel analysis ( ̃100 μl).
The clarified lysate was run over HisTrap HP columns (GE Healthcare) using a gradient from 0-100% Ni buffer B. Samples were then analyzed by SDS-PAGE gel (4-12% gel NUPAGE, Invitrogen) according to manufacturer's directions. Desired fractions were concentrated on spin filters (Vivaspin-20, Sartoris,) and then desalted over a HiPrep 26/10 Desalting column (GE heathcare) packed with Sephadex G25 resin. The G-25 buffer consisted of 50 mM HEPES, 50 mM NaCl, and 1 mM DTT, pH 7.4. The desired sample was then purified over a HiTrap Q HP column (GE) using a gradient elution from 0% Q seph buffer A to 100% Q seph buffer B (Q seph buffer A: 50 mM Tris, 0.05 M NaCl, 1 mM DTT, pH 7.6 and Q seph buffer B: 50 mM Tris, 1.0 M NaCl, 1 mM DTT, pH 7.6). Fractions containing the desired protein were analyzed and concentrated. Sample buffer was then exchanged into 50 mM HEPES, 50 mM NaCL, pH 7.4 with 1 mM DTT by passing the sample over a Hi Prep 26/10 Desalting column (GE heathcare) packed with Sephadex G25 resin. A final polishing step of Gel filtration was used when necessary. The sample was passed through a Hi Load 26/60 Superdex 200 prep grade (GE) in gel fitration buffer: (50 mM HEPES, 150 mM NaCl, 1 mM DTT, pH 7.4). Fractions were then analyzed and concentrated. The samples were then stored at −80° C. For preparation for analysis of the band, the sample is run on an SDS-PAGE gel (4-12% NUPAGE gel, Invitrogen), stained and the desired band excised and processed as described below.
III. Mass Spectrometry of Isoprene Synthase
Sample Preparation
An In-Gel Digestion and LCQ-Deca Mass Spectrometry Procedure was utilized (Modified Rosenfeld in-gel Digest Protocol) (Rosenfeld et al, Anal. Biochem. (1992) 203, 173-179; Hellman et al, Anal Biochem, (1995) 224, 451-455). The purified sample of Isoprene synthase was run on a 4-12% SDS-PAGE (NUPAGE, Invitrogen) and stained with Coomassie Brilliant Blue R-250 (Thermo Scientific) or SimplyBlue Safe Stain (Invitrogen). Band(s) of interest were excised from the gel and destained. Each gel slice was diced into small pieces  ̃1 mm×1 mm and placed into 0.65 mL “slick” (siliconized) tubes from PGC Scientific. Approximately 100 μL of 25 mM NH4HCO3/50% ACN/H2O was added to each tube and vortexed for 10 min. Supernatants were extracted and discarded. These steps were repeated twice. Then gel pieces were then run in a Savant SpeedVac to dryness ( ̃20 to 30 min).
Samples were then reducted and alkylated. For reduction, 25 μL (or enough to cover) of 10 mM DTT in 25 mM NH4HCO3 (prepared fresh) were added to dried gels. Tubes were then vortexed and spun briefly. Reactions were incubated at 50° C. for 1 hour. For alkylation, supernatants were removed and 25 μL or more of 55 mM iodoacetamide (IAA) in 25 mM NH4HCO3 were added to the gel. Reaction tubes were vortexed and spun briefly again. Reactions were allowed in dark for 1 hour at room temperature. Supernatants were removed and gels were washed with  ̃100 μL of 25 mM NH4HCO3/50% ACN/H2O, by vortexing for 10 min and briefly spinning. Supernatant were removed and the wash step was repeated once. Gel pieces were then dried in a SpeedVac ( ̃15-30 min).
Digestion buffer was prepared by adding 400 μL of 0.1% n-octyl B-D-glucopyranosidase water to 100 uL of 8M Urea. 400 uL of this digestion buffer was added to 20 ug of freshly prepared Trypsin. 0.05 μg/μL of sequencing-grade Trypsin was prepared from one vial of 20 μg sequencing grade trypsin (Promega) that was dissolved into 400 uL of 1.6 M Urea solution. Trypsin enzyme solution was added enough to cover gel pieces. Tubes were covered with parafilm and incubated at 37° C. overnight (16-20 hrs). It was ensured that there is a little extra buffer above the gel.
Peptides were extracted from gels by briefly vortexing and spinning the digest. The digest solution was transferred with gel loading tips into a 0.65 mL siliconized tube. 50 μL (enough to cover) of 50% ACN/0.1% FA/H2O were added to the gel pieces and samples were vortexed for 10 min, spun, and then sonicated for five min. Extracted peptides were pooled together in one tube. Extraction steps were repeated two to three more times until the gel pieces became white in appearance and shrank in size. Extracted digests were vortexed, spun and dried in a Speed Vac to a volume of 55 μL. In cases where the volume was less than 55 μL, enough 0.1% FA was added to make up a final volume of 55 μL.
Mass Spectrometry
The sample was injected onto a Thermofinnigan (San Jose, Calif.) LCQ-Deca electrospray ionization (ESI) ion-trap mass spectrometer. A Vydac C18 column (5μ, 300A, 0.2×150 mm, Michrom Bioresources, Auburn, Calif.) was used with a flow rate of 200 μL/min. The injection volume was 50 uL, and was filtered through an on-line trapping cartridge (Peptide CapTrap, Michrom Bioresources, Auburn, Calif.) before loading onto the column. Separation of the in-gel digest was performed with the following gradient (Solvent A: 0.1% trifluoroacetic acid in H2O (J. T. Baker, Phillipsburg, N.J.), Solvent B: 0.08% trifluoroacetic acid in acetonitrile (J. T. Baker, Phillipsburg, N.J.)):







TABLE 11-1







Gradient Table











min
A %
B %
















0
0.00
100
0



1
10.00
86
14



2
16.00
81
19



3
20.00
78
22



4
21.00
77
23



5
22.00
75
25



6
24.00
73
27



7
32.00
69
31



8
34.00
66
34



9
37.00
64
36



10
47.00
60
40



11
50.00
30
70



12
55.00
100
0



13
60.00
100
0



14
65.00
100
0



15

100
0










Mass Spectrometry Results
An aliquot of 6.4 μg of protein was loaded into 5 lanes on a 4-12% bis-tris NUPAGE gel (Invitrogen), MOPS buffer, 50 min run. As described above, the gel was stained for 2 minutes, and then de-stained for 15 minutes. The gel was washed in H2O, then all bands were excised, cut into small pieces, and destained. Gel pieces were reduced and alkyated with DTT/IAA for 1 hour each at 52° C. and RT, respectively. Trypsin was added for an overnight digestion. Extracted peptides were run on the LCQ-Deca. FIGS. 21 and 22 show the results for the mass spectrometry analysis. The lower doublet band (in FIG. 21) is identified as IspS.
FIG. 22 shows that N-terminal truncations were observed after amino acids 39, 40, 42, and 44 (according to the peptide sequence of the N-terminally His-tagged IspS protein in pDu27). The C-terminus of IspS is intact.
IV. Construction of N-Terminally Truncated IspS Variants:
All truncated constructs without affinity tags were generated using the QuickChange Site-directed Mutagenesis kit (Stratagene) using the template P. alba pET24a for PCR amplification. Approximately 50 ng of template DNA was used for amplification (with an Eppendorf Mastercycler Gradient PCR Machine) of the mutagenized PCR product with the Forward (For) and Reverse (Rev) primer pairs that correspond to each relevant truncation (QC MSV For and QC MSV Rev, for example, see Table 11-2). The following PCR reaction mixtures were used: 1 μl P. alba pET24a, 5 μl 10× PfuUltra HF buffer, 1 μl dNTP's, 1 ul (50 μM) primer-For (e.g. QC MSV For), 1 μl (50 μM) primer-Rev (e.g. QC MSV Rev), 2 μl DMSO, 39 μl diH2O, 1 μl PfuUltra HF Polymerase (Stratagene). The following PCR cycling parameters were used: 95° C. 1 min, 95° C. 30 sec., 55° C. 1 min., 68° C. 7.3 min. for one cycle followed by 95° C. 30 sec., 55° C. 1 min., 68° C. 7.3 min for a total of 18 cycles and then followed by 4° C.
The PCR products were treated with 1-2 μl of DpnI (Roche) for 1-3 hour at 37° C. A 5 μl aliquot of the DpnI treated products was visualized on a 0.8% E-gel (Invitrogen). A 1 μl aliquot of each product was transformed into chemically competent E. coli Top10 cells (Invitrogen) according to the manufacturer's protocol. Transformants were selected for on LB medium containing kanamycin at a concentration of 50 μg/ml (Kan50), and incubated overnight at 37° C. Five colonies of each transformation were selected and grown to stationary phase in 3 ml liquid LB Kan50. Plasmids were purified using a Qiagen miniprep kit according to the manufacturer's recommended protocol. Purified plasmids were sequenced (by Quintara Biosciences) with T7 Forward and Reverse primers, and confirmed for their respective truncation. The resulting plasmids (pDU39 through pDU43, see Table 11-4, FIGS. 23-34) were transformed into chemically competent E. coli BL21(λDE3)pLysS (Invitrogen) according to the manufacturer's recommended protocol. Table X describes the strains used for expression of truncated IspS enzymes.
Constructs with affinity (6×His) and proteolysis (TEV, Tobacco Etch Virus) tags were generated using P. alba pET24a as a template for PCR reactions. PCR reaction mixtures were prepared as follows: 1 ul (P. alba pET24a), 5 ul 10× PfuUltraII Fusion buffer, 1 ul dNTP's (10 mM), 1 ul primer (50 μM) Alba FL-NdeI-For or Alba TRC (MEA)-NdeI-F, 1 ul primer (50 uM) Alba FLTRC (+) TEV-R, 41 ul diH2O and 1 ul of PfuUltra II Fusion DNA Polymerase from Stratagene. PCR cycling parameters were as follows: 95° C. 1 min., 95° C. 30 sec., 55° C. 20 sec., 72° C. 25 sec. for one cycle and then repeating 95° C. 30 sec., 55° C. 20 sec., 72° C. 25 sec. for an additional 28 cycles, followed by 72° C. 3 min and then 4° C. After amplification and verification of the correct molecular weight of the product by visualization on 0.8% E-gel (Invitrogen), PCR products were digested with restriction enzymes NdeI and XhoI (Roche) for 2 hours at 37° C., and then gel purified using the Qiaquick Gel Purification system (Qiagen) according to the manufacturer's recommended protocol. 3 μl of purified product was ligated to pET-24a (Invitrogen) that was digested with NdeI and XhoI (Roche), gel purified and dephosphorylated (using SAP, shrimp alkaline phosphatase) (Promega) according to the manufacturer's recommended protocols. T4 ligase (NEB) was used for the ligation reaction, which was incubated overnight at 16° C. The ligation reaction was dialyzed into water for 30 min., and 2 μl of the reaction were used to electroporate MCM331 (see below) competent cells. Cells were allowed to recover at 30° C. for 2 hours, and then selected on Kan50 with 5 mM (R)-(−)-Mevalonolactone (MVA) (Sigma) spread onto the plate. Positive transformants were inoculated into 3 ml of liquid LB Kan50, and plasmids were isolated using the QIAPrep Spin miniprep kit (Qiagen). Inserts were verified by restriction digestion using NdeI and XhoI (Roche) and positive clones were sequenced (Quintara Biosciences) with T7 promoter and T7 terminator sequencing primers. 1 μl of each plasmid (see Table 11-4 for plasmid description and FIGS. 35-39) was transformed into chemically competent E. coli BL21(λDE3) pLysS (Invitrogen) according to the manufacturer's recommended protocol. Transformants were selected on LB Kan50+Cm35 (Chloramphenicol 35 ug/ml) plates and incubated at 37° C. See Table 11-5 for a description of all expression strains.
Strain MCM331 was prepared as follows. A synthetic operon containing mevalonate kinase, mevalonate phosphate kinase, mevalonate pyrophosphate decarboxylase, and the IPP isomerase was integrated into the chromosome of E. coli. If desired, expression may be altered by integrating different promoters 5′ of the operon.
i) Target Vector Construction
The attTn7 site was selected for integration. Regions of homology upstream (attTn7 up) (primers MCM78 and MCM79) and downstream (attTn7 down) (primers MCM88 and MCM89) were amplified by PCR from MG1655 cells. A 50 μL reaction with 1 μL 10 μM primers, 3 μL ddH2O, 45 μL Invitrogen Platinum PCR Supermix High Fidelity, and a scraped colony of MG1655 was denatured for 2:00 at 94° C., cycled 25 times (2:00 at 94° C., 0:30 at 50° C., and 1:00 at 68° C.), extended for 7:00 at 72° C., and cooled to 4° C. This resulting DNA was cloned into pCR2.1 (Invitrogen) according to the manufacturer's instructions, resulting in plasmids MCM278 (attTn7 up) and MCM252 (attTn7 down). The 832 bp ApaI-PvuI fragment digested and gel purified from MCM252 was cloned into ApaI-PvuI digested and gel purified plasmid pR6K, creating plasmid MCM276. The 825 bp PstI-NotI fragment digested and gel purified from MCM278 was cloned into PstI-NotI digested and gel purified MCM276, creating plasmid MCM281.
ii) Cloning of Lower Pathway and Promoter
MVK-PMK-MVD-IDI genes were amplified from pTrcKKDyIkIS with primers MCM104 and MCM105 using Roche Expand Long PCR System according to the manufacturer's instructions. This product was digested with NotI and ApaI and cloned into MCM281 which had been digested with NotI and ApaI and gel purified. Primers MCM120 and MCM127 were used to amplify CMR cassette from the GeneBridges FRT-gb2-Cm-FRT template DNA using Stratagene Pfu Ultra II. A PCR program of denaturing at 95° C. for 4:00, 5 cycles of 95° C. for 0:20, 55° C. for 0:20, 72° C. for 2:00, 25 cycles of 95° C. for 0:20, 58° C. for 0:20, 72° C. for 2:00, 72° C. for 10:00, and then cooling to 4° C. was used with four 50 uL PCR reactions containing 1 uL ̃10 ng/μL template, 1 μL each primer, 1.25 μL 10 mM dNTPs, 5 μL 10× buffer, 1 μL enzyme, and 39.75 μL ddH2O. Reactions were pooled, purified on a Qiagen PCR cleanup column, and used to electroporate water-washed Pir1 cells containing plasmid MCM296. Electroporation was carried out in 2 mM cuvettes at 2.5V and 200 ohms. Electroporation reactions were recovered in LB for 3 hr at 30° C. Transformant MCM330 was selected on LA with CMPS, Kan50.
iii) Integration into E. coli Chromosome
Miniprepped DNA (Qiaquick Spin kit) from MCM330 was digested with SnaBI and used to electroporate BL21(DE3) (Novagen) or MG1655 containing GeneBridges plasmid pRedET Carb. Cells were grown at 30° C. to  ̃OD1 then induced with 0.4% L-arabinose at 37° C. for 1.5 hours. These cells were washed three times in 4 C ddH2O before electroporation with 2 μL of DNA. Integrants were selected on L agar with containing chloramphenicol (5 μg/ml) and subsequently confirmed to not grow on L agar+Kanamycin (50 μg/ml). BL21 integrant MCM331 and MG1655 integrant MCM333 were frozen.







TABLE 11-2 





Primers
















MCM219
caccatgcgttgtagcgtgtcca (SEQ ID NO: 33)


 


MCM182
gggcccgtttaaactttaactagactctgcagttagcgttcaaacggcagaa



(SEQ ID NO: 34)


 


QC MSV For
gaaggagatatacatatgagcgtgtccaccgaaaatg (SEQ ID NO: 35)


 


QC MSV Rev
cattttcggtggacacgctcatatgtatatctccttc (SEQ ID NO: 36)


 


QC MVS For
gaaggagatatacatatggtgtccaccgaaaatgtgtc (SEQ ID NO: 37)


 


QC MVS Rev
gacacattttcggtggacaccatatgtatatctccttc (SEQ ID NO: 38)


 


QC MTE For
gaaggagatatacatatgaccgaaaatgtgtctttcac (SEQ ID NO: 39)


 


QC MTE Rev
gtgaaagacacattttcggtcatatgtatatctccttc (SEQ ID NO: 40)


 


QC MNV For
gaaggagatatacatatgaatgtgtctttcaccgaaac (SEQ ID NO: 41)


 


QC MNV Rev
gtttcggtgaaagacacattcatatgtatatctccttc (SEQ ID NO: 42)


 


QC MEA For
gaaggagatatacatatggaagctcgtcgttctgcg (SEQ ID NO: 43)


 


QC MEA Rev
cgcagaacgacgagcttccatatgtatatctccttc (SEQ ID NO: 44)


 


Alba
gaaggagatatacatatgcgttgtagcgtg (SEQ ID NO: 45)


FL-NdeI-For



 


Alba FLTRC (+)
cccgcgcttactcgaggccctgaaaatacaggttttcgcg


TEV-R
ttcaaacggcagaatcggtt (SEQ ID NO: 46)


 


Alba TRC (MEA)-
gaaactgaaacccatatggaagctcgtcgttctgc (SEQ ID NO: 47)


NdeI-F
















TABLE 11-3 





Primers for construction of strain MCM331

















MCM78
attTn7 up rev for
gcatgctcgagcggccgcTTTTAATCAAACATCCTGCCAACTC



integration construct
(SEQ ID NO: 48)


 


MCM79
attTn7 down rev for
gatcgaagggcgatcgTGTCACAGTCTGGCGAAACCG



integration construct
(SEQ ID NO: 49)


 


MCM88
attTn7 up forw for
ctgaattctgcagatatcTGTTTTTCCACTCTTCGTTCACTTT



integration construct
(SEQ ID NO: 50)


 


MCM89
attTn7 down forw for
tctagagggcccAAGAAAAATGCCCCGCTTACG



integration construct
(SEQ ID NO: 51)


 


MCM104
GI1.2 promoter - MVK
Gatcgcggccgcgcccttgacgatgccacatcctgagcaa




Ataattcaaccactaattgtgagcggataacacaaggagg




Aaacagctatgtcattaccgttcttaacttc




(SEQ ID NO: 52)


 


MCM105
aspA terminator - yIDI
Gatcgggccccaagaaaaaaggcacgtcatctgacgtgcc




Ttttttatttgtagacgcgttgttatagcattcta




(SEQ ID NO: 53)


 


MCM120
Forward of attTn7: attTn7
aaagtagccgaagatgacggtttgtcacatggagtt



homology, GB marker
ggcaggatgtttgattaaaagcAATTAACCCTCACTA



homology
AAGGGCGG




(SEQ ID NO: 54)


 


MCM127
Rev complement of 1.2 GI:
AGAGTGTTCACCAAAAATAATAACCTTTCCCGGTGCAgaa



GB marker homology (extra
Gttaagaacggtaatgacatagctgtttcctccttgtgtt



long), promoter, RBS, ATG
Atccgctcacaattagtggttgaattatttgctcaggatg




tggcatcgtcaagggcTAATACGACTCACTATAGGGCTCG




(SEQ ID NO: 55)
















TABLE 11-4





Plasmids for expression of IspS variants
















MD09-161
pET24a-P. alba FL C-Term (+) TEV, His tag/MCM331


MD09-163
pET24a-P. alba TRC (MEA) C-Term (+) TEV,



His tag/MCM331


pDu27
Alba-FL-pET200/D-TOPO


pDu39
Mtg pET24a-P. alba-MEA/Top10 (Untagged)


pDu40
Mtg pET24a-P. alba-MNV/Top10 (Untagged)


pDu41
Mtg pET24a-P. alba-MSV/Top10 (Untagged)


pDu42
Mtg pET24a-P. alba-MTE/Top10 (Untagged)


pDu43
Mtg pET24a-P. alba-MVS/Top10 (Untagged)
















TABLE 11-5





Strains for expression of IspS variants
















MD08-99
Alba-FL-pET200/D-TOPO (pDu27) in BL21 (λDE3) pLysS


MD09-165
BL21(λDE3)pLysS, pET24a-P. alba FL C-Term (+)



TEV, His tag


MD09-167
BL21(λDE3) pLysS, pET24a-P. alba TRC (MEA)



C-Term (+) TEV, His tag


MD09-173
BL21(λDE3)pLysS, pET24a-P. alba (MEA) Untagged



(pDu39)


MD09-174
BL21(λDE3)pLysS, pET24a-P. alba (MNV) Untagged



(pDu40)


MD09-175
BL21(λDE3)pLysS, pET24a-P. alba (MSV) Untagged



(pDu41)


MD09-176
BL21(λDE3)pLysS, pET24a-P. alba (MTE) Untagged



(pDu42)


MD09-177
BL21(λDE3)pLysS, pET24a-P. alba (MVS) Untagged



(pDu43)









V. Biochemical Analysis of IspS Truncations
The relative activity of the various N-terminally truncated IspS enzymes was determined by DMAPP assay. The strains described above were analyzed via DMAPP assay in a 96-well plate. All strains were assayed in quadruplicate. The “Full Length” variant refers to the IspS enzyme expressed in BL21(λDE3) pLysS (Invitrogen) from the P. alba pET24a plasmid.







TABLE 11-6







DMAPP Assay of N-terminal Truncations









Activity Data
Avg
OD












Variant
Average
Std Dev
% CV
OD600
Normalized















MD09-173
1125.6
93.7
8.3
5.2
217.6


MD09-174
118.6
8.0
6.7
5.2
22.8


MD09-175
1064.6
71.7
6.7
4.9
219.0


MD09-176
1179.1
64.7
5.5
4.9
238.7


MD09-177
831.7
89.6
10.8
4.9
168.2


Full Length
805.8
65.1
8.1
5.0
161.3





Results:


Table X shows that when normalized for OD600, strains MD09-173 (with plasmid pDu39), MD09-175 (pDu41), MD09-176 (pDu42), and MD09-177 (pDu43) all displayed higher DMAPP activity than the “Full Length” wild type IspS enzyme (in strain BL21(λDE3) pLysS with P. alba pET24a).






VI. Detailed Kinetic Analysis of the “MEA” Truncation in IspS
The relative specific activity was determined and the kinetics of “N-terminally truncated” P. alba isoprene synthases were examined compared to “Full length” P. alba isoprene synthases. Four strains expressing four different constructs were used in this analysis: BL21(λDE3) pLysS with P. alba pET24a; MD09-173; MD09-165; and MD09-167 (Strains described above in detail). These strains express “full length” P. alba IspS, “truncated” P. alba IspS (the MEA truncation), “full length” C-terminally TEV and His-tagged P. alba IspS, and “truncated” C-terminally TEV and His-tagged P. alba IspS, respectively. In the experiments described below, “truncated” refers specifically to the MEA variant of P. alba IspS.
All strains were inoculated into LB containing 30 mg/L chloramphenicol(Cm) and 50 mg/L kanamycin and grown overnight in 2 mL culture tubes at 37° C. The overnight cultures were diluted 1:100 in 25 mL of LB broth containing 30 mg/L chloramphenicol(Cm) and 50 mg/L kanamycin the following morning and grown at 37° C. until OD ̃0.5. Each strain was grown in triplicate. The cultures were then induced with 400 uM IPTG and incubated at 30° C. for 4 hours. 20 mL of each culture were centrifuged at 3000×g for 20 min. and the supernatant was discarded. The pellets were frozen at −80° C. overnight. Pellets were resuspended in 2 mL of a buffer containing 100 mM Tris, 100 mM NaCl, 0.25 mg/mL lysozyme and 0.25 mg/mL DNAase, pH 8. Cell suspensions were french pressed at 20,000 psi twice and the lysate was then centrifuged at 14000×g for 20 minutes to yield cell free extract that was used for kinetic studies and protein concentration determination.
To measure specific activity, 5 μL of cell free extract from each strain was incubated with 5 mM DMAPP, 50 mM MgCl2 in a buffer containing 100 mM Tris and 100 mM NaCl (pH 8) to a final volume of 100 μL for 15 min. at 30° C. in gas tight 2 mL vials. Reactions were terminated with the addition of 100 μL of 500 uM EDTA, pH 8. Samples were analyzed by GC-MS to determine the concentration of isoprene in the headspace of the vials.
To determine kcat and KM, 5 uL of cell free extract from each strain was incubated with DMAPP at concentrations ranging from 0.625 to 40 mM DMAPP in a buffer containing 100 mM Tris, 50 mM MgCl2 and 100 mM NaCl (pH 8) to a final volume of 100 μL for 15 min. at 30° C. in gas tight 2 mL vials. Reactions were terminated with the addition of 100 μL of 500 mM EDTA, pH 8. Samples were analyzed by GC-MS to determine the concentration of isoprene in the headspace of the vials. Data were analyzed using Kaleidagraph and fit to following equation for uncompetitive substrate inhibition: rate/E=kcat*S/(KM+S*(1+S/Ki)). All data were run in triplicate with the exception of MD09-167 with 2.5 mM DMAPP which was run in duplicate.
Cell free extract was run on a Caliper microfluidic electrophoresis instrument (Caliper Life Sciences, Hopkinton, Mass., USA) in order to quantify the amount of isoprene synthase in each sample. The microfluidic chip and protein samples were prepared according to the manufacturer's instructions (LabChip® HT Protein Express, P/N 760301). Culture lysates were prepared in 96-well mictrotiter plates by adding 50 mM Tris pH 8.0 containing 0.1% Tween 20, 0.1 mg/ml lysozyme, 1.0 ug/ml DNAse at room temperature for 30 minutes, followed by centrifugation. Supernatants were then transferred to another 96 well plate and stored at −20° C. until use, when they were thawed at room temperature for 30 minutes. After shaking briefly, the 2 μl of each culture sample was transferred to a 96-well PCR plate (Bio-Rad, Hercules, Calif., USA) containing 7 μl samples buffer (Caliper) followed by heating the plate to 90° C. for 5 minutes on a thermostatically controlled plate heater. The plate was allowed to cool before adding 35 μl water to each sample. The plate was then placed in the instrument along with a protein standard supplied and calibrated by the manufacturer. The instrument functions by mixing the sample with a fluorescent dye that attaches non-covalently to the proteins, followed by electrophoresis through a gel matrix. As the proteins move past a focal point in the chip, the fluorescence signal is recorded and the protein concentration is determined by quantitating the signal relative to the signal generated by a calibrated set of protein standards.







TABLE 11-7







kcat and KM and specific activity values for isoprene synthases











Isoprene
kcat ± S.D.
KM ±
Ki ±
S.A.


Synthase
(s−1)
S.D. (mM)
S.D. (mM)
(nmol/mg/min)





Full Length
0.72 ± 0.09
2.4 ± 0.3
15.7 ± 0.2 
420 ± 60


Truncated
1.5 ± 0.2
1.8 ± 0.2
9.8 ± 0.9
 800 ± 100


MD09-165
0.8 ± 0.1
2.4 ± 0.5
19 ± 4 
440 ± 80


MD09-167
1.1 ± 0.3
2 ± 1
8.7 ± 0.9
610 ± 60









Parameters were determined by fitting the following uncompetitive substrate inhibition equation to data obtained for rate/[E] vs. [DMAPP]:




  
    rate
    
      [
      E
      ]
    
  
  =
  
    
      
        k
        cat
      
      *
      
        [
        S
        ]
      
    
    
      
        K
        M
      
      +
      
        
          [
          S
          ]
        
        *
        
          (
          
            (
            
              1
              +
              
                
                  [
                  S
                  ]
                
                
                  K
                  i
                
              
            
            )
          
        
      
    
  




The specific activities (S.A.) were calculated for reactions containing 5 mM DMAPP, 50 mM MgCl2, 100 mM Tris, 100 mM NaCl, and 2.5-4.5 μg isoprene synthase from the supernatant of whole cell lysate. Reactions were performed at 30° C. for 15 minutes in triplicate using independently grown cultures.
Results/Discussion:
The specific activity of each protein was determined (FIG. 40 and Table 11-7). The specific activity of truncated isoprene synthase was approximately 2-fold greater than the specific activity of full length isoprene synthase. The C-terminally His-tagged full length isoprene synthase yielded approximately the same specific activity as the full length isoprene synthase. The C-terminally His-tagged truncated isoprene synthase gave a specific activity that was less than the non-tagged truncated isoprene synthase, but greater than the specific activity of both full length isoprene synthases.
The rate of conversion of DMAPP to isoprene was analyzed over a range of DMAPP concentrations in order to determine the kcat and KM parameters of the enzymes (FIGS. 41 and 42 and Table 11-7). The enzymes all exhibited rate profiles consistent with uncompetitive substrate inhibition by DMAPP. The Ki for the full length constructs was greater than the Ki for truncated constructs as determined by altering Ki and observing the best fit (R-value) to the data (data not shown). All data has been fit to a Ki of 10 mM in the figures in this document. The KMs of truncated isoprene synthase decreased relative to the full length isoprene synthases. Therefore, as the substrate concentration decreases the ratio between the isoprene synthase activity of the truncated isoprene synthase to the full length isoprene synthase will increase (FIG. 43). The kcats, of truncated isoprene synthases increased relative to the full length isoprene synthases. This results in greater isoprene synthase activity for the truncated isoprene synthases than the full length isoprene synthases at all substrate concentrations (FIG. 43). The ratio of the isoprene synthase activity of the truncated isoprene synthase to the full length isoprene synthase at varying DMAPP levels was determined (FIG. 43).
Conclusions: “Truncated” isoprene synthases exhibit increased kcat values and decreased KM values with respect to the catalysis of the conversion of DMAPP to isoprene compared to “full length” isoprene synthases. The specific activity of “truncated” isoprene synthases is increased relative to the specific activity of “full length” isoprene synthases. The most active isoprene synthase was non-tagged truncated isoprene synthase “MEA” variant (in strain MD09-173). The truncated isoprene synthase may increase the isoprene synthase activity in organisms used for production of isoprene compared to the full length isoprene synthase.
EXAMPLE 12
Isoprene Synthase Enrichment-Relief of DMAPP Toxicity
This example identifies residue changes within IspS that confer better activity to the enzyme through kinetic improvement, increased expression, increased solubility, or any other means by which DMAPP is more effectively converted to isoprene by Isoprene Synthase. This procedure allows for the relief of cytotoxic intracellular levels of DMAPP by expression of improved IspS variants. In a population of cells expressing a heterogeneous mixture of IspS variants, the best enzymes should allow for better growth of their host strain, and should be enriched in the mixed population.
I. Strain Construction
Construction of Strains MCM518-521 and 528-531 in which Lambda promoters drive integrated mKKDyI was as follows. Primers MCM120 and MCM224 (Table 12-1) were used to amplify the resistance cassette from the GeneBridges FRT-gb2-Cm-FRT template using Stratagene Herculase II Fusion kit according to the manufacturer's protocol. Four 50 μL reactions were cycled as follows: 95° C., 2 min; (95° C., 20 sec, 55° C., 20 sec, 72° C., 1 min) for 30 cycles; 72° C., 3 min; and 4° C. until cool. The four reactions were pooled and purified on a Qiagen PCR column according to the manufacturer's protocol and eluted with 60 μL EB at 55° C.
Plasmid pRedET-carb (GeneBridges) was electroporated into MCM446. Transformants were recovered by shaking for one hour in SOC (Invitrogen) at 30° C. and then selected on LB containing carbenicillin (50 μg/ml) (carb50) plates at 30° C. overnight. A carbenicillin resistant colony was frozen as MCM508 (Table 12-2).
Strain MCM508 was grown from a fresh streak in 5 mL LB/carb50 at 30° C. to an OD600 of  ̃0.5. 40 mM L-arabinose was added and culture was incubated at 37° C. for 1.5 hrs. Cells were harvested and electroporated with 3 μL of purified amplicons as previously, and then recovered in 500 μL SOC at 37° C. for 1.5-3 hrs. Transformants were selected on LB/kan10 plates at 37° C.
Recombination of the amplicon at the target locus was confirmed by PCR with primers GB-DW and MCM208. The resulting amplicons were sequenced to identify four clones with the sequences below. Carbenicillin-sensitive clones were frozen as strains MCM518-521.
MCM518-521 were restreaked on LB kan10 plates and grown overnight at 37° C.
Strains MCM518-521 were cultured in LB/kan10 at 37° C. and then electrotransformed with plasmid pCP20 (Datsenko K A, Wanner B L. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA. 2000 Jun. 6; 97(12):6640-5). Cells were recovered in 500 μL SOC, shaking at 30° C. for 1 hour. Transformants were selected on LB/carb50 plates at 30° C. overnight. The following morning a colony from each transformation was grown at 30° C. in liquid LB/carb50 until visibly turbid. The culture was then shifted to 37° C. for at least 3 hrs. Cells were streaked from this culture onto LB plates and grown overnight at 37° C.
The following day colonies were patched to LB, LB/carb50 and LB/kan10. Clones that grew on neither carb50 nor kan10 and were cultured in liquid LB from the patch on LB and frozen as MCM528-531.
DNA Sequences
These assemblies include the new promoters inserted on the chromosome in strains MCM518-521, as well as the very beginning of the mMVK ORF. Upstream of these assemblies is sequence from the GeneBridges FRT-gb2-Cm-FRT cassette. Downstream is the remainder of the mMVK ORF and then the rest of the lower MVA pathway integron from strain MCM508.








MCM518



(SEQ ID NO: 56)



aaagaccgaccaagcgacgtctgagagctccctggcgaattcggtaccaataaaagagctttat



 


tttcatgatctgtgtgttggtttttgtgtgcggcgcggaagttcctattctctagaaagtatag


 


gaacttcctcgagccctatagtgagtcgtattaaattcatataaaaaacatacagataaccatc


 


tgcggtgataaattatctctggcggtgttgacataaataccactggcggtgatactgagcacat


 


cagcaggacgcactgaccaccatgaaggtgcaaaggaggtaaaaaaacatggtatcctgttctg


 


cgccgggtaagatttacctgttcggtgaacacgccgtagtttatggcgaaactgcaattgcgtg


 


tgcggtggaactgcgtacccgtgttcgcgcggaactcaatgactctatcactattcagagc


 


MCM519


(SEQ ID NO: 57)



aaagaccgaccaagcgacgtctgagagctccctggcgaattcggtaccaataaaagagctttat



 


tttcatgatctgtgtgttggtttttgtgtgcggcgcggaagttcctattctctagaaagtatag


 


gaacttcctcgagccctatagtgagtcgtattaaattcatataaaaaacatacagataaccatc


 


tgcggtgataaattatctctggcggtgttgacctaaataccactggcggtgatactgagcacat


 


cagcaggacgcactgaccaccatgaaggtgcaaaggaggtaaaaaaacatggtatcctgttctg


 


cgccgggtaagatttacctgttcggtgaacacgccgtagtttatggcgaaactgcaattgcgtg


 


tgcggtggaactgcgtacccgtgttcgcgcggaactcaatgactctatcactattcagagc


 


MCM520


(SEQ ID NO: 58)



aaagaccgaccaagcgacgtctgagagctccctggcgaattcggtaccaataaaagagctttat



 


tttcatgatctgtgtgttggtttttgtgtgcggcgcggaagttcctattctctagaaagtatag


 


gaacttcctcgagccctatagtgagtcgtattaaattcatataaaaaacatacagataaccatc


 


tgcggtgataaattatctctggcggtgttgacctaaataccactggcggtgatactgagcacat


 


cagcaggacgcactgaccaccatgaaggtgcaaaggtaaaaaaacatggtatcctgttctgcgc


 


cgggtaagatttacctgttcggtgaacacgccgtagtttatggcgaaactgcaattgcgtgtgc


 


ggtggaactgcgtacccgtgttcgcgcggaactcaatgactctatcactattcagagc


 


MCM521 (in strains MCM531 and MD09-171)


(SEQ ID NO: 59)



aaagaccgaccaagcgacgtctgagagctccctggcgaattcggtaccaataaaagagctttat



 


tttcatgatctgtgtgttggtttttgtgtgcggcgcggaagttcctattctctagaaagtatag


 


gaacttcctcgagccctatagtgagtcgtattaaattcatataaaaaacatacagataaccatc


 


tgcggtgataaattatctctggcggtgttgacgtaaataccactggcggtgatactgagcacat


 


cagcaggacgcactgaccaccatgaaggtgcaaaggaggtaaaaaaacatggtatcctgttctg


 


cgccgggtaagatttacctgttcggtgaacacgccgtagtttatggcgaaactgcaattgcgtg


 


tgcggtggaactgcgtacccgtgttcgcgcggaactcaatgactctatcactattcagagc






The neo-PL.2-mKKDyI (from MCM521) was transduced into BL21(λDE3) to generate strain MD09-171. A P1 lysate of MCM521 was made and transduced into BL21(λDE3) according to standard molecular biology techniques (Miller, A Short Course in Bacterial Genetics). Transductants were selected on Kan20 LB medium plates. Positive colonies were further verified by PCR to confirm the presence of PL.2-mKKDyI in the BL21(λDE3) strain. 1 μl of pCP20 plasmid was then transformed into this strain and selected for on LB+Carb50 and incubated at 30° C. Positive transformants were subsequently streaked on an LB plate and incubated at 37° C. for loss of the pCP20 plasmid. To confirm the loss of the neomycin (kanamycin) resistance marker, colonies that grew at 37° C. were patched onto LB Kan20, LB Carb50, and plain LB plates. The strains with integrated PL.2 mKKDyI without the kanamycin resistance marker that have lost pCP20 should be sensitive to kanamycin and carbenicillin. 4 KanS CarbS were used to check by PCR for the presence of mKKDyI in BL21(λDE3) with the parental BL21(λDE3) strain as a control. Once PCR confirmed the presence of mKKDyI, the resulting strain was transformed with 1 μl of the pLysS plasmid (Invitrogen). The resulting strain, MD09-171, was used for the enrichment experiments described below.







TABLE 12-1 





Primers used for strain construction
















MCM120
aaagtagccgaagatgacggtttgtcacatggagttggcag



gatgtttgattaaaagcaattaaccctcactaaagggcgg



(SEQ ID NO: 60)


 


MCM208
gctctgaatagtgatagagtca (SEQ ID NO: 61)


 


MCM224
taaatcttacccggcgcagaacaggataccatgttttttta



cctcctttgcaccttcatggtggtcagtgcgtcctgctgat



gtgctcagtatcaccgccagtggtatttangtcaacaccgc



cagagataatttatcaccgcagatggttatctgtatgtttt



ttatatgaatttaatacgactcactatagggctcg



(SEQ ID NO: 62)


 


GB-DW
aaagaccgaccaagcgacgtctga (SEQ ID NO: 63)


 


MCM161
caccatggtatcctgttctgcg (SEQ ID NO: 64)


 


MCM162
ttaatctactttcagaccttgc (SEQ ID NO: 65)


 


MCM143
aggaggtggtctcaaatgactgccgacaacaatagta



(SEQ ID NO: 66)


 


MCM144
aggaggtggtctcagcgctctgcagttatagcattctatga



atttgcctg (SEQ ID NO: 67)
















TABLE 12-2







Strains









Strain
Description
Parent





MCM508
BL21 gi1.6-mKKDyI + predet.-carb
MCM446


MCM518
BL21 neo-PL.6-mKKDyI, clone10
MCM508


MCM519
BL21 neo-PL.0-mKKDyI, clone11
MCM508


MCM520
BL21 neo-PL.0-mKKDyI (bad RBS in front of
MCM508



mMVK), clone13


MCM521
BL21 neo-PL.2-mKKDyI, clone15
MCM508


MCM528
BL21 PL.6-mKKDyI, loopedout
MCM518


MCM529
BL21 PL.0-mKKDyI, loopedout
MCM519


MCM530
BL21 PL.0-mKKDyI (bad RBS in front of
MCM520



mMVK), loopedout


MCM531
BL21 PL.2-mKKDyI, loopedout
MCM521


MD09-171
BL21 (λDE3) PL.2-mKKDyI, loopedout +
MCM521



pLysS









II. Growth Inhibition of MCM531 by Mevalonic Acid
An overnight culture of MCM531 (see strain description) was back-diluted to an OD600 of 0.05 (this corresponds to an OD600 of approx 0.005 in a 96-well plate reader) (SpectraMax M2, Molecular Devices). The diluted culture was then aliquotted into separate wells in a 96-well deep-well plate into standard TM3 medium (13.6 g K2PO4, 13.6 g KH2PO4, 2.0 g MgSO4*7H2O) supplemented with 1% glucose and 0.8 g/L Biospringer yeast extract (1% Yeast extract final)) with 0, 1, 5, 10, 15 or 20 mM MVA added. FIG. 44 shows the growth curve of MCM531 in the various concentrations of MVA. Each MVA concentration was assayed in quadruplicate, error bars were negligible. FIG. 44 shows that MCM531 was severely inhibited for growth at concentrations of 5 mM MVA and higher.
III. Mutagenesis of IspS and Selection/Enrichment Assay
To generate a randomly mutagenized IspS open reading frame, the GeneMorph II EZ Clone domain mutagenesis kit (Stratagene) was used according to the manufacturer's recommended protocol. Specific primers to amplify the template (Pdu39 (pET24a-P. alba (MEA))) are described below (Table 12-3, pET24 Megaprime Forward and Reverse). To achieve the desired mutation frequency, the protocol outlined in the GeneMorph II kit was followed. To generate 2 to 3 residue changes per molecule, approximately 150 ng of starting template DNA was used for the initial PCR reaction. More or less template was used to generate IspS enzymes with either fewer or more residue changes, respectively. The resulting mutant “megaprimers” were then used to amplify the rest of the plasmid according to the manufacturer's recommended protocol.
The final PCR product from the GeneMorph II kit was treated with DpnI according to the manufacturer's recommended protocol. Prior to transformation into E. coli, it was necessary to desalt the PCR reaction by microdialysis. Typically, approximately 20 μl of the PCR reaction was subjected to microdialysis and used for electroporation into strain MD09-171 (Table 12-2) by standard molecular biology procedures. After electroporation, cells were recovered for 2 hours at 30° C., and then plated onto LB medium Kan50 Cm35. The entire transformation volume was plated to recover all mutations generated by the mutagenesis procedure.
For enrichment, all transformants were scraped and pooled together. An aliquot from each pool was frozen for storage in the −80. Prior to the enrichment, strains (control or experimental pools) were inoculated directly into LB with Kan50 and grown for a few hours, to minimize the loss of pool heterogeneity. After this recovery period, cells were diluted into liquid TM3 medium (described above) with Kan50, 5 mM MVA, and 200 μM IPTG. (The exact dilution was determined empirically for each pool/source of medium/control reaction). Cultures were then placed in the shaking incubator at 34° C. until an OD600 of no more than 5. At this point, plasmids were purified from 1 ml of the “enriched” culture via miniprep protocol (Qiagen). This plasmid preparation was then transformed into electrocompetent MD09-171 cells as described above. The transformed cells were recovered for 2 hours in LB medium without antibiotics, and then subjected to a subsequent round of enrichment by dilution into TM3 with Kan50, 5 mM MVA and 200 μM IPTG as described above. This culture was placed into the shaking incubator at 34° C. until it reached an OD600 of no more than 5, as described above. Plasmids were then purified, retransformed and subjected to further rounds of “enrichment.” The enrichment process continued for 5 or 6 rounds of selection, plasmid purification, and retransformation. The process continued until the culture was homogeneous, i.e. contained only one variant of IspS by sequencing analysis.
After the last round of enrichment, the plasmid pool was transformed into chemically competent E. coli Top10 cells (Invitrogen) per the manufacturer's recommended protocol, recovered, plated onto LB medium with Kan50, and sent for complete sequencing (Quintara Biosciences) for comparison to the wild type sequence of P. alba IspS. Primers used for sequencing are described below.







TABLE 12-3 





Primers used for IspS mutagenesis and sequencing
















pET24 Megaprime
gtttaactttaagaaggagatatacat


Forward



 


pET24 Megaprime 
gagctcgaattcggatcctta


Reverse



 


alba sequencing
ctcgtacaggctcaggatag


reverse



 


alba sequencing
ttacgtcccaacgctcaact


reverse2



 


EWL1000
gcactgtctttccgtctgctgc


 


QB1493
cttcggcaacgcatggaaat









IV. Individual Residue Changes Identified by Enrichment/DMAPP Toxicity Relief:
Plasmids isolated from selection/enrichment were fully sequenced. The following residue changes were confirmed by sequencing (Quintara Biosciences). The residue numbering corresponds to the P. alba “Full Length” sequence (in P. alba pET24a; SEQ ID NO:120), where the starting methionine is amino acid number 1. Identified residue changes included: V10M, F12S, T15A, E18G, V581, V58F, L70Q, L70V, L70T, T71P, V79L, E89D, G94A, S119F, F120L, G127R, E175V, T2121, S257A, R262G, A266G, F280L, N297K, F305L, L319M, E323K, A328T, D342E, A359T, K366N, E368D, L374M, S396T, V4185, K438N, H440R, T4421, T442A, I449V, A469S, K500R, K505Q, G507S, S509N, F511Y, and N532K.
Combinations of Residue Changes (in a single IspS enzyme) Identified by Enrichment/DMAPP Toxicity Relief: G127R/F511Y, L70Q/G94A/R262G/F305L, F12S/T15A/E18G/N297K, S396T/T442I, V10M/E323K, F120L/A266G, K438N/K500R, V79L/S509N, E175V/S257A/E368D/A469S, T71P/L374M, F280L/H440R, E89D/H440R, V58F/A328T/N532K, S119F/D342E/I449V, and K366N/G507S.
EXAMPLE 13
Construction of Site Saturation Libraries (SSLs) and Biochemical Analysis of the L70R Variant
This example includes an examination of possible amino acid substitutions at sites identified by the selection/enrichment procedure described above and other sites of potential interest (active site, conserved between Poplar species) for their effect on solubility, expression, and activity of IspS.
I. Strain Construction
Residues identified by the selection/enrichment procedure (L70, G94, R262, F305) described above were chosen for analysis. In addition, residues that are putatively involved in substrate binding (F303, V3065, F385, S412, Q416, F450), and residues that are different between the various Poplar species (e.g. V418, T442) were chosen for further analysis (numbering corresponds to the “full length” amino acid sequence of P. alba IspS). To generate a randomized pool of amino acid substitutions (the Site Saturation Library, SSL), pDu39 (see description above) was subjected to QuickChange (Stratagene) mutagenesis with the QC primers indicated below (Table 13-1), according to the manufacturer's recommended protocol. The PCR Reaction was prepared as follows:
1 μl pDu39, 5 μl 10× PfuUltra HF buffer, 1 μl dNTP's, 1 μl (50 μM) primer-For (e.g. QC L69 F), 1 μl (50 uM) primer-Rev (e.g. QC L69 R), 2 μl DMSO, 39 μl diH2O, and 1 μl PfuUltra HF Polymerase (Stratagene). The PCR cycling parameters for QuickChange were as follows: 95° C. 1 min, 95° C. 30 sec., 55° C. 1 min., 68° C. 7.3 min. for one cycle followed by 95° C. 30 sec., 55° C. 1 min., 68° C. 7.3 min for 17 more cycles. The temperature was then reduced to 4° C. Incorporation of the bases NNK at the codon for the residues described above allows for the insertion of codons that represent all 20 possible amino acids at the given site. 1 μl of the resulting pools of mutagenized PCR products was DpnI treated (described above), and transformed into chemically competent E. coli Top10 cells (Invitrogen) according to the manufacturer's recommended protocol. Entire transformation reactions were recovered in 1 ml LB medium without antibiotics at 37° C. for 1 hour and plated onto LB Kan50. The next day, all transformants were scraped off of the LB plates, mixed thoroughly, and plasmids were purified via miniprep (Qiagen). Pools of plasmids were then transformed into chemically competent BL21(λDE3)pLysS cells (Invitrogen) according to the manufacturer's protocol. The transformation reactions were recovered in 1 ml LB medium at 37° C. for 1 hour and then plated onto LB Kan50 Cm35 at dilutions sufficient to generate separation of positive colonies. After overnight incubation at 37° C., individual colonies were inoculated into individual wells in a 96-well deep-well microtiter plate (VWR) containing 500 ul of liquid LB Kan50 Cm35 each. In eight wells (typically column 12, A through H) strain MD09-173 (see above) or BL21(λDE3)pLysS with P. alba pET24a (Full Length) was inoculated as a control for the DMAPP assay. The microtiter plates were then sealed with a semi-permeable membrane (Breathe-Easier, Diversified Biotech), and incubated overnight at 30° C. in a shaking incubator (Vertiga). The next day, 100 μl samples from each well within a 96-well plate were mixed with 50 μl of 50% glycerol in a new 200 ul 96-well plate, and frozen at −80° C. until further analysis. This plate was then used for the DMAPP assay described below.







TABLE 13-1 





Primers used for mutagenesis
















QC L70 F
gaaaaagcagaatttnnkaccctgctggaactg (SEQ ID NO: 68)


 


QC L70 R
cagttccagcagggtmnnaaattctgctttttc (SEQ ID NO: 69)


 


QC G94 F
gagtctgatatccgtnnkgcgctggatcgcttc (SEQ ID NO: 70)


 


QC G94 R
gaagcgatccagcgcmnnacggatatcagactc (SEQ ID NO: 71)


 


QC R262 F
tcccgttggtggcgtnnkgtgggtctggcgacc (SEQ ID NO: 72)


 


QC R262 R
ggtcgccagacccacmnnacgccaccaacggga (SEQ ID NO: 73)


 


QC F303 F
tccgtcgcaaaaatgnnktctttcgtaaccatt (SEQ ID NO: 74)


 


QC F303 R
aatggttacgaaagamnncatttttgcgacgga (SEQ ID NO: 75)


 


QC F305 F
gcaaaaatgttttctnnkgtaaccattatcgac (SEQ ID NO: 76)


 


QC F305 R
gtcgataatggttacmnnagaaaacatttttgc (SEQ ID NO: 77)


 


QC V306 F
aaaatgttttctttcnnkaccattatcgacgat (SEQ ID NO: 78)


 


QC V306 R
atcgtcgataatggtmnngaaagaaaacatttt (SEQ ID NO: 79)


 


QC F385 F
gacctgtgcaacgctnnkctgcaagaagccaag (SEQ ID NO: 80)


 


QC F385 R
cttggcttcttgcagmnnagcgttgcacaggtc (SEQ ID NO: 81)


 


QC S412 F
gcatggaaatcctctnnkggcccgctgcaactg (SEQ ID NO: 82)


 


QC S412 R
cagttgcagcgggccmnnagaggatttccatgc (SEQ ID NO: 83)


 


QC Q416 F
tcttctggcccgctgnnkctggtgttcgcttac (SEQ ID NO: 84)


 


QC Q416 R
gtaagcgaacaccagmnncagcgggccagaaga (SEQ ID NO: 85)


 


QC V418 F
ggcccgctgcaactgnnkttcgcttacttcgct (SEQ ID NO: 86)


 


QC V418 R
agcgaagtaagcgaamnncagttgcagcgggcc (SEQ ID NO: 87)


 


QC T442 F
caaaaataccatgacnnkatctctcgtccttcc (SEQ ID NO: 88)


 


QC T442 R
ggaaggacgagagatmnngtcatggtatttttg (SEQ ID NO: 89)


 


QC F450 F
cgtccttcccatatcnnkcgtctgtgcaatgac (SEQ ID NO: 90)


 


QC F450 R
gtcattgcacagacgmnngatatgggaaggacg (SEQ ID NO: 91)









II. Generation of a “Winner” Plate for Secondary Assay and Identification of L70R as a Variant with Increased Specific Activity

Variants that displayed increased specific isoprene production when compared to wild type were chosen for further analysis. FIG. 45 shows a typical data set of an SSL plate for an individual residue, in this case L70. From this particular plate, the samples in wells C3 (27), D3 (39), and E3 (51) were chosen for further analysis. Other variants at different residues (listed above) that showed increased isoprene productivity, when subjected to DMAPP analysis, were isolated from their original SSL plates stored at −80° C. (described above), and re-arrayed onto a new “winner” plate for secondary screening. Two wells containing MD09-173 were included as controls. All variants were sequenced (Quintara Biosciences) and subjected to the DMAPP assay as described above. See Table 13-2 for sequencing results. For the DMAPP assay, a single growth of each variant was assayed, and therefore a single lysate generated, but in quadruplicate to generate statistically significant data. Samples were assayed at the OD600 indicated in Table 13-2. Protein analysis was performed on all lysates using the Western Breeze Western blot kit (Invitrogen) followed by fluorescence detection on a Storm860 (see below)
III. DMAPP Assay—Growth, Lysis and Isoprene Measurement
A patch plate was prepared from a glycerol stock plate using a VP-Scientific Replication Tool patch LB Agar CM35/Kan50 large patch plate from overnight glycerol stock Plate. Cultures were incubated at 30° C. overnight (20 to 24 hr). Plates were stored at 4° C. for up to a week.
An overnight growth plate was prepared from 500 mL of LB CM35/Kan50 media. 300 μL/well of LB CM35/Kan50 media were dispensed into deep 96 well plate. Using the V&S Replication Tool, the patch inoculum was transferred to a deep 96 well plate. Media was inoculated by dipping the tool then shaking the pin within the well. The overnight growth plate was sealed with a Breathe-Easier Sealing Membrane. The plate was incubated at 30° C. overnight in a Vertiga Shaking Incubator at 800 rpm for 16 to 18 hours.
A deep 96 well day growth plate was prepared by dispensing 588 μL/well of LB CM35/Kan50 media. The overnight growth plate was removed from the incubator and cultures were diluted 50-fold. 12 μL of overnight oulture was transferred to a day growth plate containing 588 μL/well of supplemented LB media. The overnight growth plate was sealed with a new Breathe-Easier Sealing Membrane and was incubated at 34° C. and 800 rpm for 2.25 hr in the Vertiga Shaking Incubator.
To induce the expression of IspS, thawed 12 mM IPTG was poured into 50 mL or 100 mL sterile reservoir and dispensed 20 μL/well into each 600 μL/well culture. Overnight growth plate was resealed with Breathe-Easier Sealing Membrane and incubated at 34° C. and 800 rpm for 4 hours in the Vertiga Shaking Incubator.
To harvest cell, 200 μL of induced culture was transferred to 450 μL Nunc storage plate. The plate was centrifuged at 3300 rpm for 20 min at 4° C. in a low speed benchtop centrifuge. 180 μL supernatant was removed with a pipettor and discarded. The plate was sealed with an aluminum foil membrane, covered with a plastic plate lid and stored frozen at −80° C.
The OD600 of the plates were read. 150 μL 1×PBS was dispensed into a 96 well Costar Read Plate (#9017). 50 μL of culture sample was then transferred to the read plate. The OD600 reading was then taked with a Spectramax Plate Reader.
Lysis: The harvest plate was defrosted in a room temperature water bath for 4 min and then incubated in Thermomixer at 25° C. at 1200 rpm for 1 min. Lysis buffer was dispensed at 80 μL/well to 20 μL/well of harvest cells. 1.25× Lysis Working Stock buffer was prepared from 6.25 ml 1M Tris pH 8, 625 μl 10% Tween 20, 312.5 μl 0.2 M PMSF, 462.5 μl 10 mg/ml DNAse I (Sigma), 1.25 ml 1 M MgCl2, 132.5 μl 25000 U/μl Lysozyme (Epicentre Technologies) and 40.968 ml dIH20. Plates were incubated on a Thermomixer at 25° C. and 1200 rpm for 30 min. 1× Lysis Buffer stock was prepared by diluting 19 ml of 1.25× Lysis Working stock with 4.75 ml dIH2O.
DMAPP Working Stock was prepared as follows.










Dimethylallyl Pyrophosphate (triammonium salt) 25 mg (Cayman Chemicals,


Cat No. 63180)















final
total vol
diluent
stock vol
stock
dilution
Reagent
Units
Plate(s)


















30
840
840
solid
25000
1
DMAPP
mg/mL
1


3
2500
2250
250
30
10
DMAPP
mg/mL
1









The Diluent was 0.1 M Potassium Phosphate. 200 uL/well was dispensed for transfer to sample wells and was stored on ice.
DMAPP Reaction: 1× Lysis Buffer was dispensed at 65 μL/well. 15 μL/well of lysate was transferred to the respective sample wells in a 96 Deep well Zinsser Glass Block. DMAPP reagent was dispensed at 20 μL/well. The glass block was sealed with an aluminum foil membrane and incubated at 25° C. at 450 rpm for 45 min. The reaction was stopped by transferring the Glass Block to a 70° C. water bath and incubating for 6 min. GC Analysis was performed as previously described.
IV. Western Blot of Isoprene Synthase with Fluorescence Labeled Secondary Antibody.
Samples were prepared and run on NativePAGETM Novex® Bis-Tris Gels (Invitrogen) according to the manufacturer's protocol. After completion of the run the gels were immediately transferred to Nitrocellulose membranes using the XCell IITM Blot Module (Invitrogen) according to the manufacturer's recommended protocol. After transfer, the membrane was placed in 15 ml of the appropriate Blocking Solution (Ultra filtered Water 31.5 ml, Blocker/Diluent (Part A) 9 ml, Blocker/Diluent (Part B) 4.5 ml) in the covered, plastic dish provided in the kit and incubated for 30 minutes on a rotary shaker set at 1 revolution/sec. The Blocking Solution was decanted and the membrane rinsed 2 times with 20 ml of water for 5 minutes. The membrane was incubated with 15 ml of Primary Antibody (Ab) Solution (24 μl of primary Ab in 15 ml Blocking solution) for 1 hour, followed by washing 3 times 5 minutes with 20 ml of 1× Antibody Wash Solution. The membrane was then incubated in 15 ml of SecondaryAntibody Solution (15 ul secondary Ab (Alexa Fluor 488 goat anti-rabbit IgG (H+L, Invitrogen)) in 15 of blocking solution) for 30 minutes. The membrane was washed 3 times at 5 minutes with 20 ml of Antibody Wash, and rinsed two times 2 minutes with 20 ml of water. The membrane was dried between paper towels and stored at room temperature for further detection. The fluorescent bands were detected and quantified using the Storm 860 Molecular Imager (GMI, Inc).
V. Results:
Table 13-2 shows all of the relevant data for each variant assayed: sequencing results, residue change, average isoprene production, protein concentration, and average specific activity (of all 4 replicates). FIG. 46 shows the graphical representation of the data shown in Table 13-2. Specific activity was calculated by multiplying the isoprene produced (μg/l) by 0.0414 and then dividing by protein concentration (mg/ml). This conversion factor (0.0414) accounts for the total headspace volume in a sealed 2 ml GC vial (1.9 ml), the lysate volume (15 ul), the duration of the DMAPP assay (45 min), and the molecular weight of isoprene. Thus, specific activity values are given in nmol isoprene/mg Isps/min.
The data in Table 13-2 and the graph in FIG. 46 show that of all variants analyzed, all three L70R variants displayed higher specific activity than wild type. To analyze the L70R variants further, the specific activity values for all 3 isolates (4 replicates of each) were averaged and compared to the controls (2 isolates, 4 replicates each). Therefore, there were 12 measurements for the L70R variant, and 8 for wild type. These data are shown below in Table 13-3. When corrected for protein, the L70R variants displayed a 25% increase in activity over MD09-173 (the MEA truncation). FIG. 47 shows the average specific activity for all L70R variants compared to MEA. Error bars show one standard deviation. The two data sets for L70R and the MEA control were subjected to a Student's T-Test for statistical analysis, which yielded a P-value of 6.0011×10−5.







TABLE 13-2





Sequencing Results, Isoprene Production, Protein concentration,


and Specific Activity for all residues in the Winner Plate






























Note that sequence of all plasmids is identical to Pdu39 (see above) with the exception of the indicated codon. The L70R variants are highlighted in gray.













TABLE 13-3







Average Specific Activity of all L70R variants relative to the MEA


control.











Variant Average
Specific Activity
Standard Deviation















L70R
365.116
35.31977



WT
294.7809
17.19228



(the MEA control)







See FIG. 47 for bar graph.






EXAMPLE 14
Truncations of P. alba, P. tremuloides, P. trichocharpa, and Kudzu Isoprene Synthases
This example describes the generation of a series of truncations in the IspS enzymes of P. alba, P. tremuloides, P. trichocharpa, and Kudzu and to determine their effect on activity.
I. Strain Construction
All isoprene synthase genes were codon optimized for E. coli, synthesized, and cloned into pET24a by DNA2.0 (Menlo Park, Calif.). All truncated constructs were generated using the QuickChange Site-Directed Mutagensis kit (Stratagene) using the previously described templates P. alba pET24a (for plasmids pDu47-3 through -7, FIGS. 48, 49, 51-60), P. tremuloides pET24a (plasmid pDu48, FIGS. 49C, 61 and 62), P. trichocharpa pET24a (pDu49, FIGS. 50A, 63, 64), or pET24d-Kudzu (pDu50 and 50-4, FIGS. 50B, 50C, 65-68) for PCR amplification. Approximately 50 ng of template DNA was used for amplification (with an Eppendorf Mastercycler Gradient PCR Machine) of the PCR product with the Forward (For) and Reverse (Rev) primer pairs that correspond to each relevant truncation (QC Trunc −3 F and QC Trunc −3 R, for example, see Table 14-1). PCR reactions mixtures were as follows: 1 μl P. alba pET24a (or other template), 5 μl 10× PfuUltra HF buffer, 1 μl dNTP's (10 mM), 1 μl (50 uM) primer-For, 1 μl (50 μM) primer-Rev, 1.5 μl DMSO, 39.5 μl diH2O and 1 μl PfuUltra HF Polymerase. PCR cycle parameters were as follows: (95° C. 1 min., 95° C. 1 min., 55° C. 1 min., 68° C. 7.30 min.) for 18 cycles then 4° C. until cool using an Eppendorf Mastercycler Gradient Machine. The PCR products were treated with 1-2 μl of DpnI (Roche) for 1-3 hour at 37° C. 5 μl of the DpnI treated products were visualized on a 0.8% E-gel (Invitrogen). 1 μl of each product was transformed into chemically competent E. coli Top10 cells (Invitrogen) (according to the manufacturer's protocol). Transformants were selected for on LB medium containing kanamycin at a concentration of 50 μg/ml (Kan50), and incubated overnight at 37° C. Five colonies of each transformation were selected and grown to stationary phase in 3 ml liquid LB Kan50. Plasmids were purified using a QIAPrep Spin miniprep kit (Qiagen) according to the manufacturer's recommended protocol. Purified plasmids were sequenced (by Quintara Biosciences) with T7 Forward and Reverse primers, compared to the parental sequence, and confirmed for their respective truncation. The resulting plasmids (pDu47-3 through pDu50-4, see Table 14-2) were transformed into chemically competent E. coli BL21(DE3) pLysS (Invitrogen) according to the manufacturer's recommended protocol. Table 14-3 describes the strains used for expression of truncated IspS enzymes.
After overnight incubation at 37° C., individual colonies were inoculated into individual wells in a 96-well deep-well microtiter plate (VWR) containing 500 μl of liquid LB Kan50 CM35 each. Microtiter plates were then sealed with a semi-permeable membrane (Breathe-Easier, Diversified Biotech), and incubated overnight at 30° C. in a shaking incubator (Vertiga). The next day, 100 μl samples from each well within a 96-well plate were mixed with 50 μl of 50% glycerol in a new 200 μl 96-well plate, and frozen at −80° C. until further analysis. This plate was then used for the DMAPP assay as described in Example 13. Table 14-4 shows the average specific productivity of all samples, and FIG. 69 shows the graphical representation of the same data.
DMAPP activity and protein quantitation was determined as described in Example 13.
Specific activity was calculated by multiplying the isoprene produced (μg/1) by 0.00776 and then dividing by protein concentration (mg/ml). This conversion factor (0.00776) accounts for the total headspace volume in a sealed 2 ml GC vial (1.9 ml), the lysate volume (80 μl), the duration of the DMAPP assay (45 min), and the molecular weight of isoprene. Thus, specific activity values are given in nmol isoprene/mg IspS/min.







TABLE 14-1 





Primers
















QC Trunc-3 F
gaaggagatatacatatgaccgaagctcgtcgt (SEQ ID NO: 92)


 


QC Trunc-3 R
acgacgagcttcggtcatatgtatatctccttc (SEQ ID NO: 93)


 


QC Trunc-4 F
gaaggagatatacatatggaaaccgaagctcgt (SEQ ID NO: 94)


 


QC Trunc-4 R
acgagcttcggtttccatatgtatatctccttc (SEQ ID NO: 95)


 


QC Trunc-5 F
gaaggagatatacatatgactgaaaccgaagct (SEQ ID NO: 96)


 


QC Trunc-5 R
agcttcggtttcagtcatatgtatatctccttc (SEQ ID NO: 97)


 


QC Trunc-6 F
gaaggagatatacatatggaaactgaaaccgaa (SEQ ID NO: 98)


 


QC Trunc-6 R
ttcggtttcagtttccatatgtatatctccttc (SEQ ID NO: 99)


 


QC Trunc-7 F
gaaggagatatacatatgaccgaaactgaaacc (SEQ ID NO: 100)


 


QC Trunc-7 F
ggtttcagtttcggtcatatgtatatctccttc (SEQ ID NO: 101)


 


QC Kudzu MEA F
agaaggagatataccatggaagctcgtcgttccgcaaac (SEQ ID NO: 102)


 


QC Kudzu MEA R
gtttgcggaacgacgagcttccatggtatatctccttct (SEQ ID NO: 103)


 


QC Kudzu-4 F
agaaggagatataccatggagcataattcccgt (SEQ ID NO: 104)


 


QC Kudzu-4 R
acgggaattatgctccatggtatatctccttct (SEQ ID NO: 105)


 


QC Trem/Trich-2 F
gaaggagatatacatatggaaacgcgtcgttct (SEQ ID NO: 106)


 


QC Trem/Trich-2 R
agaacgacgcgtttccatatgtatatctccttc (SEQ ID NO: 107)
















TABLE 14-2





Plasmids


















pDu47-3
Mtg pET24a-P.alba TRC (-3)



pDu47-4
Mtg pET24a-P.alba TRC (-4)



pDu47-5
Mtg pET24a-P.alba TRC (-5)



pDu47-6
Mtg pET24a-P.alba TRC (-6)



pDu47-7
Mtg pET24a-P.alba TRC (-7)



pDu48
Mtg pET24a-P.tremu TRC (MET)



pDu49
Mtg pET24a-P.tricho TRC (MET)



pDu50
Mtg pET24d-Kudzu TRC (MEA)



pDu50-4
Mtg pET24d-Kudzu TRC (-4)

















TABLE 14-3





Strains


















MD09-197-3
BL21(DE3)pLysS, pDu47-3



MD09-197-4
BL21(DE3)pLysS, pDu47-4



MD09-197-5
BL21(DE3)pLysS, pDu47-5



MD09-197-6
BL21(DE3)pLysS, pDu47-6



MD09-197-7
BL21(DE3)pLysS, pDu47-7



MD09-198
BL21(DE3)pLysS, pDu48



MD09-199
BL21(DE3)pLysS, pDu49



MD09-200
BL21(DE3)pLysS, pDu50



MD09-200-4
BL21(DE3)pLysS, pDu50-4

















TABLE 14-4







Specific Productivity of Variants listed above









Isoprene Produced (μg/l)
OD600
OD














Sample
Sample
Average
Std Dev
% CV
Raw
xDF xCF
Norm

















IspS N
MD09-197-3
3381
154
5
0.3060
3.4
986.6


terminal
MD09-197-4
3000
206
7
0.3160
3.5
847.7


Truncation
MD09-197-5
2932
266
9
0.3200
3.6
818.2


Variants
MD09-197-6
2450
217
9
0.3040
3.4
719.4



MD09-197-7
2285
397
17
0.3020
3.4
675.5



MD09-198
1916
106
6
0.3330
3.7
513.6



MD09-199
2031
108
5
0.2140
2.4
847.6



MD09-200
141
8
6
0.3700
4.1
34.0



MD09-200-4
1829
197
11
0.2760
3.1
591.5



MD09-173
2414
201
8
0.3400
3.8
633.9



MD09-176
2826
354
13
0.3260
3.7
773.9



BL21 DE3 pLysS +
2175
117
5
0.2990
3.3
649.6



P alba pET24a









A second experiment was conducted with the strains outlined in Table 14-5. Control was BL21 DE3 pLysS with P. alba pET24a (full length P. alba IspS).







TABLE 14-5





Strains


















MD09-197-3
BL21(DE3)pLysS, pDu47-3



MD09-197-4
BL21(DE3)pLysS, pDu47-4



MD09-197-5
BL21(DE3)pLysS, pDu47-5



MD09-197-6
BL21(DE3)pLysS, pDu47-6



MD09-197-7
BL21(DE3)pLysS, pDu47-7



MD09-198
BL21(DE3)pLysS, pDu48



MD09-199
BL21(DE3)pLysS, pDu49



MD09-173
BL21(DE3)pLysS, pET24a-P.alba (MEA) Untagged




(pDu39)



MD09-174
BL21(DE3)pLysS, pET24a-P.alba (MNV) Untagged




(pDu40)



MD09-175
BL21(DE3)pLysS, pET24a-P.alba (MSV) Untagged




(pDu41)



MD09-176
BL21(DE3)pLysS, pET24a-P.alba (MTE) Untagged




(pDu42)



MD09-177
BL21(DE3)pLysS, pET24a-P.alba (MVS) Untagged




(pDu43)



MD09-197-3
BL21(DE3)pLysS, pDu47-3



MD09-197-4
BL21(DE3)pLysS, pDu47-4










Results

All truncations of P. alba IspS and two from P. tremuloides and P. trichocharpa were assayed in parallel to compare their relevant specific activities via DMAPP assay and quantitative Western blot. At least two samples per variant were assayed for isoprene production and amount of IspS in mg/ml. Variant MD09-174 produced little isoprene and expressed little protein, yet displayed high specific activity. High specific activities were also displayed by MD09-173, MD09-176, and MD09-197-3 (see Table 14-6 and FIG. 70). The highest levels of protein (μg) in 3 μg total protein were displayed by MD09-176 and MD09-197-3, indicating that these variants are more effectively expressed in the E. coli BL21 DE3 host strain.







TABLE 14-6







Specific Activity of truncations.











Average Specific

Average ug IspS/


Strain
Activity
Standard Deviation
3 μg total protein













P. alba FL
240.16239
31.0423851
0.198


MD09-175
331.8755329
8.958408319
0.188


MD09-177
340.1959506
39.72104203
0.150


MD09-176
363.516921
3.376026129
0.202


MD09-174
452.7792122
27.71567075
0.018


MD09-197-7
279.2042431
23.82331163
0.158


MD09-197-6
309.8357305
7.903564316
0.164


MD09-197-5
293.22592
20.97161876
0.165


MD09-197-4
321.3926574
56.13760028
0.186


MD09-197-3
333.0604155
45.92710529
0.207


MD09-173
368.4597405
37.80631246
0.159


MD09-198
297.6476631
56.81405985
0.154


MD09-199
256.7342861
8.239697653
0.216









EXAMPLE 15
Constructs for Three-Dimensional Structure Determination
I. Construction of pMAL-C4X Kudzu
A synthetic gene, coding for isoprene synthase (IspS) of the Kudzu vine (Pueraria lobata) and codon-optimized for E. coli, was purchased from DNA2.0 (Menlo Park, Calif.) and provided as plasmid p9795 (FIGS. 71 and 72). The insert was removed by digestion with BspLU11I/PstI, gel-purified, and religated into NcoI/PstI-digested pTrcHis2B (Invitrogen, Carlsbad, Calif.). The resulting plasmid was named pTrcKudzu (FIG. 73: map of pTrcKudzu). The stop codon in the insert is before the PstI site, which results in a construct in which the His-Tag is not attached to the IspS protein.
A PCR reaction was performed to amplify the E. coli codon-optimized Kudzu gene using plasmid pTrcKudzu as the DNA template, primers EL-959 and EL-960, 10 mM dNTP (Roche, Indianapolis, Ind.), and Pfu Ultra II Fusion DNA polymerase (Stratagene, La Jolla, Calif.) according to manufacturer's protocol. PCR conditions were as follows: 95° C. for 2 min (first cycle only), 95° C. for 25 sec, 60° C. for 25 sec, 72° C. for 30 sec, repeat for 28 cycles, with final extension at 72° C. for 1 min. The PCR product was then purified using the QIAquick PCR Purification Kit (Qiagen Inc, Valencia, Calif.).
The Kudzu PCR product (1 μg) was digested using EcoRI and HindIII restriction endonucleases (Roche) according to manufacturer's protocol. The digest was incubated 37° C. for 30 minutes to minimize digestion of the internal EcoRI site that is present in the Kudzu gene. The digested PCR fragment was then purified using the QIAquick PCR Purification Kit. The vector pMAL-C4X (0.5 μg) (New England Biolabs, Ipswich, Mass.; FIGS. 75 and 76) was digested using EcoRI and HindIII restriction endonucleases (Roche) according to manufacturer's protocol. The digested vector was then gel purified using the QIAquick Gel Extraction Kit (Qiagen Inc). A DNA ligation reaction was performed using T4 DNA ligase (New England Biolabs) with a 5:1 ratio of digested Kudzu PCR product to digested pMAL-C4X vector according to manufacturer's protocol. An aliquot of the ligation reaction was then transformed into TOP10 chemically competent cells (Invitrogen Corp). Transformants were selected on LA+50 μg/μl carbenicillin plates.
Screening of transformants containing the Kudzu gene was performed by picking colonies and performing PCR with primers EL-957 and EL-966 using PuReTaq Ready-To-Go PCR beads (GE Healthcare, Piscataway, N.J.) according to manufacturer's protocol. PCR conditions were as follows: 95° C. for 2 min (first cycle only), 95° C. for 30 sec, 50° C. for 30 sec, 72° C. for 40 sec, repeat for 28 cycles, with final extension at 72° C. for 1 min. PCR products were analyzed on a 2% E-gel (Invitrogen Corp) looking for a 600 bp fragment. Colonies containing the correct sized PCR product insert were submitted for DNA sequencing using primers EL-950, EL-951, EL-953, and EL-957. DNA sequencing confirmed the construction of plasmid pMAL-C4X Kudzu (FIGS. 77-79).







TABLE 15-1 







Primer sequences








Primer



name
Primer sequence





EL-950
CGGTGAACTGAAAGGTGACGTCC (SEQ ID NO: 108)


 


EL-951
GGACGTTAACGCTATTAACACCCTG (SEQ ID NO: 109)


 


EL-953
CACATCGTCGATCAGCTCCAGC (SEQ ID NO: 110)


 


EL-957
GGTCGTCAGACTGTCGATGAAGCC (SEQ ID NO: 111)


 


EL-959
GCTTATGAATTCTGTGCGACCTCTTCTCAATTTACTCAG



(SEQ ID NO: 112)


 


EL-960
GCTTATAAGCTTAGACATACATCAGCTGGTTAATCGGG



(SEQ ID NO: 113)


 


EL-966
CTCCTCCAGCAGGTTCTCACC (SEQ ID NO: 114)









Plasmid pMAL-C4X Kudzu was transformed into OneShot BL21(λDE3) chemically competent cells (Invitrogen Corp). Expression strain transformants were selected on LA+50 mg/ml carbenicillin plates.
II. IspS Variants for Crystal Structure Trials
This example describes methods to generate affinity tagged isoprene synthase (IspS) enzymes for expression, purification and crystallization.
Strain Construction
For constructs in the pET200D-TOPO vector (Invitrogen), PCR products of the IspS enzymes from P. alba, P. tremuloides, and P. trichocharpa were gel extracted and purified (Qiagen), using 0.8% E-gel (Invitrogen), according to the manufacturer's recommended protocol. PCR reactions for pET200 constructs are as follows: Reaction mixture was 1 μl (Templates)-pET24a-P.alba, 5 μl 10× PfuUltraII Fusion buffer, 1 μl dNTP's (10 mM), 1 μl primer (50 uM) primer F-(MCM219 or 218), 1 μl primer (50 uM) primer R-(MCM182), 41 μl diH2O and 1 μl of PfuUltra II Fusion DNA Polymerase from Stratagene; Cycle Parameter were 95° C. 1 min., 95° C. 1 min, 55° C. 20 sec., 72° C. 27 sec. for 29 cycles followed by 72° C. for 3 min and then 4° C. until cool, using an Eppendorf Mastercycler. Similar reactions were performed for P. tremuloides, P. trichocarpa, and Kudzu. 3 μl of purified product was then ligated to the pET200D/TOPO vector (Invitrogen), according to the manufacturer's protocol. The reaction was incubated for 5 minutes at room temperature, and the 6 μl topoisomerase mixture was then transformed into E. coli Top10 chemically competent cells (Invitrogen) according to the manufacturer's protocol. Transformants were selected for on LB Kan50, and incubated at 37° C. overnight. Five colonies per construct were chosen and screened using PuReTaq Ready-To-Go PCR Beads (Amersham) using the T7 Forward and MCM182 primers (Table 15-2). Clones harboring inserts of the correct size were further verified by plasmid miniprep using the QIAPrep Spin Miniprep kit (Qiagen) followed by sequencing using the T7 Forward and T7 Reverse primers (Quintara Biosciences). One fully sequenced construct for each IspS variant (see below for details and sequence/FIGS. 79-90), was chosen for further study. 1 μl of each plasmid was transformed into BL21(λDE3) pLysS (Invitrogen) according to the manufacturer's protocol. Transformants were selected for on LB medium with Kan50+Cm35 and incubated at 37° C. overnight. The resulting strains were used for expression and purification of various IspS enzymes for crystallography studies.
Construction of N-terminally 6His-tagged IspS plasmids, strains and purification is described in Example 11.







TABLE 15-2 





Primers
















MCM219
caccatgcgttgtagcgtgtcca (SEQ ID NO: 114)


 


MCM182
gggcccgtttaaactttaactagactctgcagttagcgttc



aaacggcagaa (SEQ ID NO: 115)


 


MCM218
caccatgcgtcgttctgcgaactac (SEQ ID NO: 116)
















TABLE 15-3





Plasmids
















P. alba pET24a
pET24a with “full length” IspS from P. alba


P. trichocharpa
pET24a with “full length” IspS from P. trichocharpa


pET24a


P. tremuloides
pET24a with “full length” IspS from P. tremuloides


pET24a


MBP-Kudzu


pDu27
P. albaFL-pET200/Top 10


pDu30
P. albaTRC-pET200/Top10


pDu31
P. tremTRC-pET200/Top 10


pDu32
P. trichTRC-pET200/Top10


MD09-161
pET24a-P.alba FL C-Term (+) TEV,



His tag/MCM331


MD09-163
pET24a-P.alba TRC (MEA) C-Term (+) TEV,



His tag/MCM331
















TABLE 15-4





Strains
















MBP-Kudzu



MD08-99
BL21 DE3 pLys + pDu27


MD08-100
BL21 DE3 pLys + pDu30


MD08-102
BL21 DE3 pLys + pDu31


MD08-104
BL21 DE3 pLys + pDu32


MD09-165
BL21(DE3)pLysS, pET24a-P.alba FL C-Term (+) TEV,



His tag


MD09-167
BL21(DE3) pLysS, pET24a-P.alba TRC (MEA) C-Term



(+) TEV, His tag









III. Digestion of TEV (Tobacco Etch Virus) or EK (Enterokinase)-Tagged Enzymes

TEV Cleavage (IspS from Strains MD09-165 and MD09-167)

Strains MD09-165 and MD09-167 are described in Example 11. For digestion, enzymes were purified through a Ni charged sepharose (GE Healthcare) and desalted into 50 mM HEPES, 50 mM NaCl pH 7.4 buffer containing 1 mM DTT. Digestion was performed with TurboTEV Protease from Eton Bioscience Inc. One unit of TurboTEV per 10 μg of purified protein was used. The digest was performed at 4° C. overnight. Samples were passed through another Ni column equilibrated in the Ni buffer to remove uncleaved enzyme, tag, TurboTEV protease (that is also tagged) and impurities. The Ni column pass though and washes were analyzed using SDS-PAGE gel (NUPAGE, Invitrogen) and DMAPP activity assays. Samples containing pure enzyme were pooled and desalted into 50 mM NaCl pH 7.4 buffer containing 1 mM DTT and stored at −80° C.
EK Cleavage (IspS from Strains MD08-102 and MD08-104)
For digestion enzymes were purified through a Ni charged sepharose (GE Healthcare) and desalted into 50 mM HEPES, 50 mM NaCl pH 7.4 buffer containing 1 mM DTT.
Digestion was performed with EKMax (E180-02) (Invitrogen) using 1 unit of EKMax per 20 μg of purified protein at 4° C. overnight. Samples were passed over EK Away resin (Invitrogen) to remove excess enterokinase. Samples were batched onto Ni charged sepharose resin (equilibrated in the Ni wash buffer) and incubated for 30 min at 4° C., with occasional inverting. This removed uncleaved enzyme, tag, and impurities. The Ni column pass through and washes were analyzed using SDS-PAGE gel (4-12% NUPAGE, Invitrogen) and DMAPP activity assays. Samples containing pure enzyme were pooled and desalted into 50 mM HEPES, 50 mM NaCl pH 7.4 buffer containing 1 mM DTT and stored at −80° C.
IV. Purification of MBP-IspS
Construction of pMAL-C4X Kudzu for the expression of MBP-Kudzu isoprene synthase is described above. MBP-Kudzu isoprene synthase production from E. coli grown in batch culture at the 15-L scale.
Medium Recipe (Per Liter Fermentation Medium)
K2HPO4 7.5 g, MgSO4*7H2O 2 g, citric acid monohydrate 2 g, ferric ammonium citrate 0.3 g, yeast extract 0.5 g, 1000× Modified Trace Metal Solution 1 ml. All of the components were added together and dissolved in diH2O. This solution was autoclaved. The pH was adjusted to 7.0 with ammonium hydroxide (30%) and q.s. to volume. Glucose 10 g, thiamine*HCl 0.1 g, and antibiotics were added after sterilization and pH adjustment.
1000× Modified Trace Metal Solution
Citric Acids*H2O 40 g, MnSO4*H2O 30 g, NaCl 10 g, FeSO4*7H2O 1 g, CoCl2*6H2O 1 g, ZnSO*7H2O 1 g, CuSO4*5H2O 100 mg, H3BO3 100 mg, NaMoO4*2H2O 100 mg. Each component is dissolved one at a time in diH2O, pH to 3.0 with HCl/NaOH, then q.s. to volume and filter sterilized with 0.22 micron filter.
Fermentation was performed in a 15-L bioreactor with BL21 (DE3) E. coli cells containing the pMAL-C4X plasmid expressing a maltose binding protein (MBP)-Kudzu isoprene synthase fusion molecule. This experiment was carried out to produce isoprene synthase at the desired fermentation pH 7.0 and temperature 30° C. A frozen vial of the E. coli strain was thawed and inoculated into tryptone-yeast extract medium. After the inoculum grew to OD 1.0, measured at 550 nm (OD550), 120 mL was used to inoculate a 15-L bioreactor bringing the initial volume to 9-L.
Expression of the desired molecule was achieved by adding isopropyl-beta-D-1-thiogalactopyranoside (IPTG). The IPTG concentration was brought to 1 mM when the OD550 reached a value of 10. Cells containing the desired product were harvested 3 hrs after IPTG addition.
MBP-IspS Purification
The broth was centrifuged for 15 min at 10000×g. The pellet was collected and frozen at −80° C. until further purification. Cells were resuspended in MBP-Bind Buffer (5% glycerol, 20 mM Tris pH 7.4, 200 mM NaCl, 2 mM DTT, 1 mg/ml lysozyme) and passed through the french press three times at 20000 psi. The lysate was then ultracentrifuged at 100000×g for 1 hour to yield a relatively clear solution. The supernatant was pipetted from the top of the tube without disturbing the gelatinous material on the bottom of the centrifuge tube. Gel filtration was performed on the supernatant using a Superdex-200 26/60 column (GE healthcare). The column was developed using MBP-Bind buffer at a flow rate of 3 mL/min at 23° C. Fractions were tested for DMAPP activity as described below. Active fractions were pooled and loaded onto 25 mL amylose resin (New England Biolabs). The column was washed with 10 column volumes MBP-Bind buffer and the protein was then eluted with 2 column volumes of MBP-Bind buffer containing 10 mM maltose to yield >90% pure MBP-IspS.
V. DMAPP Assay
The following reaction mixture was used for the DMAPP assay: 25 μL lysate mixture, 5 μL MgCl2 (1 M), 5 μL DMAPP (100 mM), and 65 μL 100 mM Tris pH 8, 100 mM NaCl for a total volume of 100 μL. The reaction is performed at 30° C. for 15 minutes in a gas tight 1.8 mL GC tube. Reactions are terminated by the addition of 100 μL 500 mM EDTA (pH 8). The amount of isoprene produced was measured by GC/MS as described above.
EXAMPLE 16
Three-Dimensional Structure of IspS
Seven constructs of plant isoprene synthase (IspS) were prepared to generate crystals suitable for x-ray diffraction. These were: a construct containing N-terminal histidine-tagged maltose binding protein and kudzu IspS (MBP-kudzu), full-length P. alba IspS with N-terminal histidine-tag (MD08-99), P. alba IspS with the first nineteen N-terminal residues removed (MD08-100), this construct also had the N-terminal his-tag removed after purification. Full-length, untagged P. alba IspS (strain RM11608-2). A truncated P. alba IspS construct featuring two additional residues before the twin-arginine motif was generated (MD09-167). P. tricharpa IspS was generated, which contains both an N-terminal his-tag and N-terminal truncation (MD08-104), and another construct composed of IspS from P. tremuloides was generated with an N-terminal his-tag and N-terminal truncation (MD08-102). Construction of strains expressing various isoprene synthases are described above.
Each construct was purified and a concentrated protein solution was then prepared for surveying possible crystallization conditions. Each construct was purified independently and surveyed as described below. All in-house crystallization screens were set up using the hanging drop vapor diffusion method. At a minimum, each construct was surveyed using the following commercial screens: the Crystal Screen from Hampton Research (Aliso Viejo, Calif.) and the JCSG+Suite from Qiagen (Valencia, Calif.).
Purified MBP-kudzu using was set up using the following commercial screens: the Crystal Screen from Hampton Research and the JCSG+Suite from. Additionally, purified MBP-kudzu was sent to the Hauptman-Woodward Institute (Buffalo, N.Y.) for high-throughput screening, where no fewer than 1536 conditions were surveyed. The purified MBP-kudzu fusion precipitated out of solution in the majority of conditions, and no protein crystals were observed.
The next construct used for crystallization screening was MD08-99 (full-length P. alba IspS with N-terminal histidine-tag). MD08-99 was purified and the histidine-tag was removed. The same three initial crystallization screens were performed as for MBP-kudzu. The Hampton Research Crystal Screen and Qiagen JCSG+Suite were each performed at multiple protein concentrations. Small needle-like crystals were observed in some Hampton Research Crystal Screen conditions. Further attempts to improve the crystals involved co-crystallization with the IspS inhibitor sodium ibandronate (Sigma-Aldrich, St Louis, Mo.). Taken together, an additional 288 crystallization conditions were attempted with variations of pH, concentration, and crystallization reagents. The nine best crystals were then prepared for data collection and tested in-house on a Rigaku RU200 rotating anode generator and R-AXIS IV++, and they either did not diffract x-rays or were salt crystals.
The first nineteen N-terminal residues of P. alba IspS were removed to produce construct MD08-100. This construct had the N-terminal histidine-tag removed after purification. In house crystallization screens were performed using the Hampton Research Crystal Screen and Qiagen JCSG+Suite, each with multiple protein concentrations. Initial crystal hits included hexagonal plates that diffracted to 16 Å resolution, and small rods that diffracted to 5 Å resolution using the in-house x-ray generator. In an attempt to improve the crystals, MD08-100 was co-crystallized with either sodium ibandronate or sodium pyrophosphate (Sigma-Aldrich, St Louis, Mo.), both of which are inhibitors of the IspS activity. Neither inhibitor resulted in improved crystals or improved diffraction. An additional 168 crystallization conditions were attempted with variations of pH, concentration, and crystallization reagents. The twenty-one most promising MD08-100 crystals were screened for diffraction, with the best resolution obtained being 5 Å.
Full-length, untagged P. alba IspS (strain RM11608-2) from a fermentation run was purified. An initial screen was set up using the Hampton Research Crystal Screen, and crystals were observed in four different conditions. All four crystals were tested for diffraction in-house, with three being salt crystals and one not diffracting.
A truncated P. alba IspS construct featuring two additional residues before the twin-arginine motif was generated (MD09-167). This construct contains a C-terminal histidine-tag, and crystallization experiments were set up with the tag either cleaved or not cleaved, at varying protein concentrations, and with or without sodium pyrophosphate. Initial crystallization screens were done as per MBP-kudzu. Crystals from this construct were observed in numerous conditions; optimization included 528 variations of pH, precipitating agents, concentrations, and inhibitors. From the optimization experiments, fifteen different MD09-167 crystals were screened in-house for diffraction. In an effort to improve the resolution, various crystal freezing conditions were tested, with the effect of improving the diffraction limits from 10 Å to 6.5 Å.
A new construct containing P. tricharpa IspS was generated, which contains both an N-terminal histidine-tag and an N-terminal truncation (MD08-104). Purified MD08-104 with cleaved histidine-tag was surveyed using the Hampton Research Crystal Screen and the Qiagen JCSG+suite. This construct generated heavier precipitate than the P. alba IspS constructs. Very small needles were observed, with none of the crystals being suitable for diffraction.
Another construct composed of IspS from P. tremuloides was generated with an N-terminal histidine-tag and an N-terminal truncation (MD08-102). Purified MD08-102 with and without cleaved histidine-tag was set up using the Hampton Research Crystal Screen and the Qiagen JCSG+Suite at varying protein concentrations. Rod and plate-like crystals were observed in some conditions and an additional 120 experiments were performed to improve the crystals by varying pH, concentration, and crystallization reagents. From the optimization experiments, ten crystals were tested in-house, with the initial best diffraction reaching 5 Å. Upon further modification of the freezing conditions of the crystals, a crystal was found that diffracted to 3.3 Å from the non-cleaved histidine-tagged protein. This crystal was grown by mixing 2 μL of protein (10 mg/ml, with 30 mM MgCl2) with 2 μL of precipitant solution [10% (wt/vol) polyethylene glycol 8000, 0.1 M HEPES, pH 7.5, 8% ethylene glycol] and equilibrated against 500 μL of precipitant. A cluster of rod-shaped crystals appeared after three weeks. The crystals belong to the tetragonal space group P43212, and have unit cell dimensions a=154.2, b=154.2, c=142.7.
In-house x-ray diffraction data were collected under a nitrogen stream at 100 K using a Rigaku RU200 generator and R-AXIS IV++ detector. Before flash-freezing the crystal in liquid nitrogen, it was cryoprotected by swiping it through a solution containing 10% (wt/vol) polyethylene glycol 8000, 0.1 M HEPES, pH 7.5, and 25% ethylene glycol. Data were integrated using Mosflm (Leslie, A. (1998) J. of Appl. Crystallography 30, 1036-1040) and scaled using SCALA (Collaborative Computational Project, N. (1994) Acta Crystallographica Section D 50, 760-763). The data were then phased by molecular replacement using MrBUMP (Keegan, R. M., and Winn, M. D. (2007) Acta Crystallographica Section D 63, 447-457; Vagin, A., and Teplyakov, A. (1997) J. of Appl. Crystallography 30, 1022-1025), with a monomer of limonene synthase (Protein Data Bank ID 2ONH)(Berman, H., et al. (2007) Nucl. Acids Res. 35, D301-303) as the starting model. The crystal contains one dimer in the asymmetric unit with a solvent content of 66%.
A 3.05 Å data set from the same crystal was then collected using beamline 11-1 of the Stanford Synchrotron Radiation Laboratory. These data were also processed using Mosflm and SCALA. Data collection and refinement statistics are given in Table 16-1.
Refinement with Refmac5 (Collaborative Computational Project, N. (1994) Acta Crystallographica Section D 50, 760-763) was used with iterative manual rebuilding steps using the visualization program Coot (Emsley, P., and Cowtan, K. (2004) Acta Crystallographica Section D 60, 2126-2132). During refinement, the geometry of the protein was checked using Molprobity (Davis, I. W., et al. (2007) Nucl. Acids Res., gkm216).
The fold of poplar IspS is similar to bornyl diphosphate synthase (Whittington, D. A., et al. (2002) Proc. Natl. Acad. Sci. USA 99, 15375-15380), limonene synthase (Hyatt, D. C., et al. (2007) Proc. Natl. Acad. Sci. USA 104, 5360-5365), and tobacco 5-epi-aristolochene synthase (Starks, C. M., et al. (1997) Science 277, 1815-1820). The structure consists of two helical domains, a C-terminal domain containing the active site and N-terminal domain (FIGS. 90 and 91). Coordinates are provided in Table 16-7.







TABLE 16-1





Data Collection and Refinement Statistics


















Data Collection




Space Group
P43212



Cell dimensions



a, b, c (Å)
154.2, 154.2, 142.7



α, β, γ, (°)
90.0, 90.0, 90.0



Resolution (Å)
37.8-3.05



Rmerge
 16.4 (72.9)a



<I/σI>
10.3 (2.6)



Completeness (%)
 99.8 (100)



Redundancy
 7.3 (7.4)



Refinement



Resolution (Å)
37.8-3.05



No. measured reflections
248741



No. Unique reflections
34201



Rwork
21.1



Rfree
27.1



rmsd bonds, (Å)
0.011



rmsd angles, (°)
1.28



No. of Atoms



Protein, ionsb
8331



Water
18










Flexible Loops

The unique and unexpected discovery coming from the determination of the three dimensional structure of isoprene synthase is that several crucial loops forming the active site are flexible. The discovery can be immediately seen when the known structure of other terpene synthases are compared with the structure of isoprene synthase (FIG. 94). Overall, the structures are highly conserved in the conformation of secondary structure and connectivity loops. (In this example of the Poplar tremuloides IspS from construct P.tremTRC-pET200, the numbering convention is such that the first number of the complete sequence containing the tag is −35, with the first residue of IspS being 1.) However, three segments, forming a considerable portion of the substrate binding pocket, notably the truncated N-terminus, along with two loops comprised of residues 438-453 (Loop I) and residues 512-527 (Loop II) are seen to diverge (FIGS. 95 to 97). This has been attributed to the absence of substrate complexed with the enzyme in our structure determination.
In comparing the enzyme with BdpS, for example, we find that the loops corresponding to residues 498-513 and 573-587 are composed of the same number of residues and have a homologous, but not identical amino sequence in these regions. We expect that the related terpene synthases will be found to display similar flexibility in the segments as these structure become more thoroughly studied. The residues in terpene synthases corresponding to these variable loop regions are enumerated in Table 16-2.







TABLE 16-2







Residues corresponding to variable loops in terpene synthases












Poplar IspS
LS
BdpS
TEAS

















N-term I
Met 1
Met 57
Ile 54
Val 14




Arg 2
Arg 58
Arg 55
Arg 15




Arg 3
Arg 59
Arg 56
Pro 16




Ser 4
Ser 60
Ser 57
Val 17




Ala 5
Gly 61
Gly 58
Ala 18




Asn 6
Asn 62
Asn 59
Asp 19




Tyr 7
Tyr 63
Tyr 60
Phe 20




Glu 8
Asn 64
Gln 61
Ser 21




Pro 9
Pro 65
Pro 62
Pro 22




Asn 10
Ser 66
Ala 63
Ser 23




Ser 11
Arg 67
Leu 64
Leu 24




Trp 12
Trp 68
Trp 65
Trp 25




Asp 13
Asp 69
Asp 66
Gly 26




Tyr 14
Val 70
Ser 67
Asp 27




Asp 15
Asn 71
Asn 68
Gln 28




Tyr 16
Phe 72
Tyr 69
Phe 29



N-term II
Leu 17
Ile 73
Ile 70
Leu 30




Leu 18
Gln 74
Gln 71
Ser 31




Ser 19
Ser 75
Ser 72
Phe 32




Ser 20
Leu 76
Leu 73
Ser 34




Asp 21
Leu 77
Asn 74
Ile 35




Thr 22
Ser 78
Thr 75
Asp 36




Asp 23
Asp 79
Pro 76
Asn 37




Glu 24
Tyr 80
Tyr 77
Gln 38




Ser 25
Lys 81
Thr 78
Val 39




Ile 26
Glu 82
Glu 79
Ala 40




Glu 27
Asp 83
Glu 80
Glu 41




Val 28
Lys 84
Arg 81
Lys 42



Loop I
Leu 438
Leu 498
Leu 498
Thr 446




Ala 439
Gly 499
Gly 499
Ala 447




Ser 440
Thr 500
Thr 500
The 448




Ala 441
Ser 501
Ser 502
Tyr 449




Ser 442
Val 502
Tyr 503
Glu 450




Ala 443
Glu 503
Phe 504
Val 451




Glu 444
Glu 504
Glu 505
Glu 452




Ile 445
Val 505
Leu 506
Lys 453




Ala 446
Ser 506
Ala 507
Ser 454




Arg 447
Arg 507
Arg 508
Arg 455




Gly 448
Gly 508
Gly 509
Gly 456




Glu 449
Asp 509
Asp 510
Gln 457




Thr 450
Val 510
Val 511
Ile 458




Ala 451
Pro 511
Pro 512
Ala 459




Asn 452
Lys 512
Lys 513
Thr 460




Ser 453
Ser 513
Thr 514
Gly 461



Loop II
Tyr 512
Tyr 576
Tyr 573
Tyr 520







Ile 521




His 513
His 577
Leu 574
His 522




Asn 514
Asn 578
His 575
Asn 523




Gly 515
Gly 579
Gly 576
Leu 524




Asp 516
Asp 580
Asp 577
Asp 525




Ala 517
Gly 581
Gly 578
Gly 526




His 518
His 582
Phe 579
Tyr 527




Thr 519
Gly 583
Gly 580
The 528




Ser 520
Thr 584
Val 581
His 529




Pro 521
Gln 585
Gln 582
Pro 530




Asp 522
His 585
His 583
Glu 531




Glu 523
Pro 586
Ser 584
Lys 532




Leu 524
Ile 587
Lys 585
Val 533




Thr 525
Ile 588
Thr 586
Lue 534




Arg 526
His 589
Tyr 587
Lys 535










This important finding can be exploited for the engineering of improved isoprene synthase in a straightforward manner. It would be desirable to exploit the flexibility to enhance enzyme performance by making substitutions in the amino aids forming these segments to facilitate the transitions the enzyme must undergo in the steps of binding substrate and allowing rearrangement of substrate in different kinetic steps that are postulated to occur during enzymatic de-phosphorylation and for electron transfer to convert DMAPP to isoprene.
The structure provides the new insight that these loops can be present in at least two conformations: the “open” form in the absence of substrate, as we have see in the uncomplexed structure of the isoprene synthase, and a “closed,” or active form when the substrate is bound. It would therefore also be beneficial to modify residues coming in contact with the loops in the active form as described in Table 16-3.







TABLE 16-3







Residues coming within 5 angstroms of flexible elements












P trem IspS
1N1B
2ONG
5EASe





N-term
L17, L18, S19, S20,
70I, 71Q, 72S, 73L,
I73, Q74, S75, L76,
L30, S31, F32, S33,


neigbors
S239, R243, F253,
298S, 302S, 312F,
F299, R303, F313,
S248, K252, Y262,



A254, R255, D256,
313V, 314R, 315D,
A314, R315, A316,
A263, R264, D265,



R257, I259, E260,
316R, 318V, 319E,
R317, V319, E320,
R266, V268, E269,



D293, Y295, D296,
352D, 354Y, 355D,
D353, Y355, D356,
D302, F304, D305,



V297, Y298, G299,
356V, 357Y, 358G,
V357, Y358, G359,
A306, Y307, G308,



T300, E303, Y325,
359T, 362E, 384Y,
T360, E363, Y385,
T309, E312, Y334,



L374, Y375, elements
433Y, 434H, elements
F434, Y435, elements
F383, I384, elements of



of loop I, elements of
of loop I, elements of
of loop I, elements of
loop I, elements of loop



loop II, V529, L530,
loop II, 589I, 590A,
loop II, M590, T591,
II, I538, I538, V543



T534
594F
F595


Loop I
Elements of N-term,
Elements of N-term,
Elements of N-term,
Elements of N-term,


neighbors
D293, Y295, V297,
352D, 354Y, 356V,
D353, Y355, V357,
D304, F304, A306,



E370, A371, W373,
429E, 430A, 432W,
E430, A431, W433,
E379, S380, W382,



L374, S378, T379,
433Y, 437Y, 438T,
F434, H438, K439,
F383, Y387, T388,



P380, F382, Y385,
439P, 441L, 444Y,
P440, L442, Y445,
P389, V391, Y394,



F386, R433, L434
445L, 493R, 494L,
L446, R493, L494,
L395, R441, V442,



C435, N436, D437,
495P, 496D, 497D,
A495, D496, D497,
I443, D444, D445,



V454, S455, C456,
514I, 515Q, 516C,
L514, Q515, C516,
I462, E463, C464,



Y457, M458, T469,
517Y, 518M, 529V,
Y517, M518, R529,
C465, M466, M477,



V472, I476, Y512,
532V, 536I, 572Y,
V532, I536, Y573,
F480, A484, Y520,



elements of loop II
elements of loop II
elements of loop II
elements of loop II


Loop II
Elements of N-
Elements of N-
Elements of N-
Elements of N-


neighbors
terminus, E187, L188,
terminus, 246D, 247L,
terminus, D247, I248,
terminus, E195, Q196,



R255, R257, F270,
314R, 316R, 329E,
R315, R317, E330,
R264, R266, F279,



E271, Q273, Y274,
330S, 332F, 333W,
P331, Q333, H334,
E280, Q282, Y283,



F285, V288, A439,
344I, 348I, 499G,
N345, I347, G499,
I294, I297, A447,



S440, S442, S508,
500T, 503Y, 568A,
T500, V502, A569,
T448, E450, V516,



H509, C510, T511,
569Q, 570F, 571I,
Q570, L571, M572,
E517, V518, T519,



Y512, R528, V529,
572Y, 588H, 589I,
Y573, Q589, M590,
Y520, H537, I538,



L530, S531, V532
590A, 591G, 592L
T591, R592, T593
I539, N540, L541









Selection of Sites for Improvement of Plant Isoprene Synthase

The isoprene synthases of plants were expected to be homologous to the terpene synthases. The three-dimensional structures of three homologous terpene synthases have been determined: Salvia officinalis bornyl diphosphate synthase (BdpS; pdb entry 1N1B), Mentha spicata limonene synthase (LS; pdb entry 2ONG), and tobacco 5-epi-aristolochene synthase (TEAS; pdb entry SEAS). These enzymes share only 33% homology but their tertiary structure is conserved. Sequence identity is shown in Table 16-4, and structural homology between the structures is shown in Table 16-5. In addition, the structures of intermediate complexes with all three related enzymes have shown that not only tertiary folding, but also detailed interactions in the active sites of these enzymes are highly conserved.







TABLE 16-4







Percent Identity of Terpene Cyclases.












P trem IspS
1N1B
2ONG
5EASe

















P alba IspSa
98.6
40.7
41.3
33.2



P trem IspSb

41.0
41.4
33.2



1N1Bc


51.4
33.8



2ONGd



33.3







aPolar alba isoprene synthase



bPolar tremuloides isoprene synthase



cbornyl diphosphate synthase



dlimonene synthase



e5-epi-aristolochene synthase













TABLE 16-5







Structural Alignment of Terpene Synthases











1N1Bb
2ONGc
5EASd
















P trem IspSa
1.40 (465)e
1.29 (468)
1.62 (458)



1N1B

1.27 (520)
1.97 (476)



2ONG


1.83 (477)







aPolar tremuloides isoprene synthase



bbornyl diphosphate synthase



climonene synthase



d5-epi-aristolochene synthase



eRoot mean square deviation in Å for Cα atoms, with the number of aligned residues in parenthesis






In this example of the Poplar tremuloides IspS from construct P.tremTRC-pET200, the numbering convention is such that the first number of the complete sequence containing the tag is −35, with the first residue of IspS being 1.
A comparison of the active site from the structure of BdpS and the structure of poplar IspS indicates that the active site involved in metal ion binding and phosphate recognition is conserved. In particular, Arg 255, Asp 292, Asp 296, Glu 370, Arg 433 and Asn 436 of poplar IspS were observed to overlap equivalent residues in BdpS. The positioning of an intermediate of the BdpS was also compared with the poplar IspS structure. Based on this it was possible to identify the analogous binding region and the approach direction that dimethylallyl pyrophosphate would require in order to bind and react with the poplar IspS enzyme.
Based on the structure of poplar IspS, sites in the poplar IspS were identified as candidates for mutagenesis to produce variant IspS enzymes with improved performance. Briefly, sites were selected in the IspS that might alter the interaction of the metal binding, diphosphate recognition, DMAPP chain binding and/or the approach to the active site.
I. Diphosphate/Metal Binding Sites
The side chains of amino acid residues in the poplar IspS that are found in proximity to the metal and diphosphate binding side chains were identified. These residues include Asp 293, Tyr 385, Ser 392, and Asp 437. Engineering of these sites may result in increased enzyme activity.
II. Substrate Access Loops
The substrate access loops of poplar IspS are in regions that deviate from the BdpS structure. In the BdpS structure the residues create a cover over the active site. It is likely that upon substrate binding the structure of poplar IspS will form a similar structure. As such the residues in these loops, including residues 440-453 and 512-524, may be in a position to alter the activity of the poplar IspS. In the poplar IspS enzyme, residues 440-453 have the sequence SASAEIARGETANS and residues 512-526 have the sequence YHNGDAHTSPDEL.
III. Isoprenyl Binding Site
The complex of BdpS and the product of the reaction, bornyl diphosphate (PDB entry 1N24), was used to identify residues in the poplar IspS structure that with protein engineering may be used modulate substrate specificity and/or reaction rate (altered on and off rates of substrate and product). These residues include Ser 261, Trp 264, Phe 285, Thr 289, Ser 393, Ser 394, Phe 432, and Try 512.







TABLE 16-6







Candidate mutagenesis sites.









Poplar IspS














DPP/Metal Binding Sites
Asp 293




Tyr 385




Ser 392




Asp 437



Substrate Access Loop I
Ser 440




Ala 441




Ser 442




Ala 443




Glu 444




Ile 445




Ala 446




Arg 447




Gly 448




Glu 449




Thr 450




Ala 451




Asn 452




Ser 453



Substate Access Loop II
Tyr 512




His 513




Asn 514




Gly 515




Asp 516




Ala 517




His 518




Thr 519




Ser 520




Pro 521




Asp 522




Glu 523




Leu 524



Isoprenyl Binding Site
Ser 261




Trp 264




Phe 285




Thr 289




Ser 393




Ser 394




Phe 432




Tyr 512

















TABLE 16-7





Coordinates of P. tremuloides IspS

















HEADER
----
XX-XXX-XX  xxxx









COMPND
---



REMARK
3


REMARK
3
REFINEMENT.


REMARK
3
 PROGRAM : REFMAC 5.5.0088


REMARK
3
 AUTHORS : MURSHUDOV, VAGIN, DODSON


REMARK
3


REMARK
3
 REFINEMENT TARGET: MAXIMUM LIKELIHOOD


REMARK
3


REMARK
3
 DATA USED IN REFINEMENT.


REMARK
3
 RESOLUTION RANGE HIGH (ANGSTROMS): 3.05


REMARK
3
 RESOLUTION RANGE LOW (ANGSTROMS): 110.17


REMARK
3
 DATA CUTOFF  (SIGMA(F)): NONE


REMARK
3
 COMPLETENESS FOR RANGE  (%): 99.67


REMARK
3
 NUMBER OF REFLECTIONS  : 32446


REMARK
3


REMARK
3
 FIT TO DATA USED IN REFINEMENT.


REMARK
3
 CROSS-VALIDATION METHOD  : THROUGHOUT


REMARK
3
 FREE R VALUE TEST SET SELECTION: RANDOM


REMARK
3
 R VALUE  (WORKING + TEST SET): .21396


REMARK
3
 R VALUE   (WORKING SET): .21092


REMARK
3
 FREE R VALUE    : .27112


REMARK
3
 FREE R VALUE TEST SET SIZE (%): 5.1


REMARK
3
 FREE R VALUE TEST SET COUNT  : 1727


REMARK
3


REMARK
3
 FIT IN THE HIGHEST RESOLUTION BIN.


REMARK
3
 TOTAL NUMBER OF BINS USED  :  20


REMARK
3
 BIN RESOLUTION RANGE HIGH  :  3.050


REMARK
3
 BIN RESOLUTION RANGE LOW  :  3.129


REMARK
3
 REFLECTION IN BIN (WORKING SET):  2359


REMARK
3
 BIN COMPLETENESS (WORKING + TEST) (%): 100.00


REMARK
3
 BIN R VALUE (WORKING SET): .288


REMARK
3
 BIN FREE R VALUE SET COUNT  : 127


REMARK
3
 BIN FREE R VALUE    :  .352


REMARK
3


REMARK
3
 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.


REMARK
3
 ALL ATOMS    :  8349


REMARK
3


REMARK
3
B VALUES.


REMARK
3
 FROM WILSON PLOT   (A**2): NULL


REMARK
3
 MEAN B VALUE  (OVERALL, A**2): 24.592


REMARK
3
 OVERALL ANISOTROPIC B VALUE.


REMARK
3
 B11 (A**2):  .41


REMARK
3
 B22 (A**2):  .41


REMARK
3
 B33 (A**2):  −.81


REMARK
3
 B12 (A**2):  .00


REMARK
3
 B13 (A**2):  .00


REMARK
3
 B23 (A**2):  .00


REMARK
3


REMARK
3
ESTIMATED OVERALL COORDINATE ERROR.


REMARK
3
 ESU BASED ON R VALUE    (A): NULL


REMARK
3
 ESU BASED ON FREE R VALUE   (A): .427


REMARK
3
 ESU BASED ON MAXIMUM LIKELIHOOD   (A): .327


REMARK
3
 ESU FOR B VALUES BASED ON MAXIMUM LIKELIHOOD (A**2): 39.836


REMARK
3


REMARK
3
CORRELATION COEFFICIENTS.


REMARK
3
 CORRELATION COEFFICIENT FO-FC  :  .916


REMARK
3
 CORRELATION COEFFICIENT FO-FC FREE:  .868


REMARK
3


REMARK
3
RMS DEVIATIONS FROM IDEAL VALUES  COUNT RMS  WEIGHT


REMARK
3
 BOND LENGTHS REFINED ATOMS  (A): 8495; .011; .022


REMARK
3
 BOND LENGTHS OTHERS  (A): 5804; .001; .020


REMARK
3
 BOND ANGLES REFINED ATOMS (DEGREES): 11476; 1.279; 1.953


REMARK
3
 BOND ANGLES OTHERS  (DEGREES): 14093; .882; 3.000


REMARK
3
 TORSION ANGLES, PERIOD 1 (DEGREES): 1020; 7.002; 5.000


REMARK
3
 TORSION ANGLES, PERIOD 2 (DEGREES): 435; 35.412; 24.299


REMARK
3
 TORSION ANGLES, PERIOD 3 (DEGREES): 1525; 18.250; 15.000


REMARK
3
 TORSION ANGLES, PERIOD 4 (DEGREES): 58; 16.811; 15.000


REMARK
3
 CHIRAL-CENTER RESTRAINTS  (A**3): 1266; .070; .200


REMARK
3
 GENERAL PLANES REFINED ATOMS  (A): 9416; .005; .020


REMARK
3
 GENERAL PLANES OTHERS  (A): 1780; .001; .020


REMARK
3


REMARK
3
ISOTROPIC THERMAL FACTOR RESTRAINTS.  COUNT RMS  WEIGHT


REMARK
3
 MAIN-CHAIN BOND REFINED ATOMS (A**2): 5104; .514; 1.500


REMARK
3
 MAIN-CHAIN BOND OTHER ATOMS (A**2): 2068; .059; 1.500


REMARK
3
 MAIN-CHAIN ANGLE REFINED ATOMS (A**2): 8204; 1.000; 2.000


REMARK
3
 SIDE-CHAIN BOND REFINED ATOMS (A**2): 3391; 1.218; 3.000


REMARK
3
 SIDE-CHAIN ANGLE REFINED ATOMS (A**2): 3272; 2.157; 4.500


REMARK
3


REMARK
3
NCS RESTRAINTS STATISTICS


REMARK
3
 NUMBER OF DIFFERENT NCS GROUPS: 1


REMARK
3


REMARK
3
NCS GROUP NUMBER   : 1


REMARK
3
 CHAIN NAMES    : A B


REMARK
3
 NUMBER OF COMPONENTS NCS GROUP: 1


REMARK
3
  COMPONENT C SSSEQI TO C SSSEQI CODE


REMARK
3
  1  A  17  A  541  6


REMARK
3
  1  B  17  B  541  6


REMARK
3
    GROUP CHAIN  COUNT RMS  WEIGHT


REMARK
3
LOOSE POSITIONAL 1 1  (A): 7038; .37; 5.00


REMARK
3
LOOSE THERMAL  1 1 (A**2): 7038; 1.09; 10.00


REMARK
3


REMARK
3
TWIN DETAILS


REMARK
3
NUMBER OF TWIN DOMAINS: NULL


REMARK
3


REMARK
3


REMARK
3
TLS DETAILS


REMARK
3
 NUMBER OF TLS GROUPS: 8


REMARK
3
 ATOM RECORD CONTAINS RESIDUAL B FACTORS ONLY


REMARK
3


REMARK
3
 TLS GROUP: 1


REMARK
3
 NUMBER OF COMPONENTS GROUP: 1


REMARK
3
 COMPONENTS  C SSSEQI TO C SSSEQI


REMARK
3
 RESIDUE RANGE: A  17  A  219


REMARK
3
 ORIGIN FOR THE GROUP (A): −64.7667 37.6643 −.0896


REMARK
3
 T TENSOR


REMARK
3
  T11:  .0648 T22:  .0357


REMARK
3
  T33:  .0787 T12:  .0200


REMARK
3
  T13:  .0129 T23:  −.0089


REMARK
3
 L TENSOR


REMARK
3
  L11:  3.7204 L22:  1.5111


REMARK
3
  L33:  2.6701 L12:  .5715


REMARK
3
  L13:  .6692 L23:  −.9699


REMARK
3
 S TENSOR


REMARK
3
  S11:  .0562 S12:  .0478 S13: −.1976


REMARK
3
  S21:  −.1702 S22:  −.0055 S23:  .1376


REMARK
3
  S31:  .0900 S32:  −.2188 S33: −.0507


REMARK
3


REMARK
3
 TLS GROUP: 2


REMARK
3
 NUMBER OF COMPONENTS GROUP: 1


REMARK
3
 COMPONENTS  C SSSEQI TO C SSSEQI


REMARK
3
 RESIDUE RANGE: A  220  A  287


REMARK
3
 ORIGIN FOR THE GROUP (A): −59.5787 8.4529 −.7693


REMARK
3
 T TENSOR


REMARK
3
  T11:  .1615 T22:  .0645


REMARK
3
  T33:  .1539 T12:  −.0314


REMARK
3
  T13:  −.0461 T23:  .0198


REMARK
3
 L TENSOR


REMARK
3
  L11:  2.4192 L22:  4.6709


REMARK
3
  L33:  .7709 L12: −3.2943


REMARK
3
  L13:  −.1814 L23:  −.0705


REMARK
3
 S TENSOR


REMARK
3
  S11:  .0055 S12:  −.0699 S13: −.2073


REMARK
3
  S21:  −.1805 S22:  .0996 S23:  .3781


REMARK
3
  S31:  .2596 S32:  .0887 S33: −.1051


REMARK
3


REMARK
3
 TLS GROUP:  3


REMARK
3
 NUMBER OF COMPONENTS GROUP: 1


REMARK
3
 COMPONENTS  C SSSEQI TO C SSSEQI


REMARK
3
 RESIDUE RANGE: A  288  A  374


REMARK
3
 ORIGIN FOR THE GROUP (A): −40.1866 1.6932 .5805


REMARK
3
 T TENSOR


REMARK
3
  T11:  .1149 T22:  .1003


REMARK
3
  T33:  .1629 T12:  .0153


REMARK
3
  T13:  .0224 T23:  .0164


REMARK
3
 L TENSOR


REMARK
3
  L11:  .2271 L22:  .7399


REMARK
3
  L33:  4.8529 L12:  .3413


REMARK
3
  L13:  .4755 L23:  −.1746


REMARK
3
 S TENSOR


REMARK
3
  S11:  −.0449 S12:  −.0288 S13: −.1131


REMARK
3
  S21:  −.1346 S22:  −.0665 S23: −.2749


REMARK
3
  S31:  −.0040 S32:  .1558 S33:  .1114


REMARK
3


REMARK
3
 TLS GROUP: 4


REMARK
3
 NUMBER OF COMPONENTS GROUP: 1


REMARK
3
 COMPONENTS  C SSSEQI TO C SSSEQI


REMARK
3
 RESIDUE RANGE: A  375  A  541


REMARK
3
 ORIGIN FOR THE GROUP (A): −47.2220 21.5399 6.9217


REMARK
3
 T TENSOR


REMARK
3
  T11:  .1551 T22:  .1194


REMARK
3
  T33:  .1485 T12:  −.0666


REMARK
3
  T13:  .0275 T23:  .0272


REMARK
3
 L TENSOR


REMARK
3
  L11:  2.2352 L22:  2.1698


REMARK
3
  L33:  2.3370 L12:  −.4501


REMARK
3
  L13:  2.2662 L23:  −.1852


REMARK
3
 S TENSOR


REMARK
3
  S11:  .0233 S12:  −.3041 S13: −.0323


REMARK
3
  S21:  .3375 S22:  .0236 S23:  .0121


REMARK
3
  S31:  .0592 S32:  −.2979 S33: −.0469


REMARK
3


REMARK
3
 TLS GROUP:  5


REMARK
3
 NUMBER OF COMPONENTS GROUP: 1


REMARK
3
 COMPONENTS  C SSSEQI TO C SSSEQI


REMARK
3
 RESIDUE RANGE: B  17  B  219


REMARK
3
 ORIGIN FOR THE GROUP (A): −73.9834 −39.9016 −18.5783


REMARK
3
 T TENSOR


REMARK
3
  T11:  .0658 T22:  .1153


REMARK
3
  T33:  .1251 T12:  −.0621


REMARK
3
  T13:  −.0164 T23:  −.0098


REMARK
3
 L TENSOR


REMARK
3
  L11:  4.6230 L22:  1.7260


REMARK
3
  L33:  3.8816 L12:  −.4202


REMARK
3
  L13: −1.8646 L23:  −.9046


REMARK
3
 S TENSOR


REMARK
3
  S11:  −.0685 S12:  .0375 S13: −.0003


REMARK
3
  S21:  .1931 S22:  .0510 S23: −.0097


REMARK
3
  S31:  .0317 S32:  −.2047 S33:  .0175


REMARK
3


REMARK
3
 TLS GROUP:  6


REMARK
3
 NUMBER OF COMPONENTS GROUP: 1


REMARK
3
 COMPONENTS  C SSSEQI TO C SSSEQI


REMARK
3
 RESIDUE RANGE: B  220  B  287


REMARK
3
 ORIGIN FOR THE GROUP (A): −62.1586 −12.7634 −18.1912


REMARK
3
 T TENSOR


REMARK
3
  T11:  .1825 T22:  .0804


REMARK
3
  T33:  .1512 T12:  .0549


REMARK
3
  T13:  .0773 T23:  .0208


REMARK
3
 L TENSOR


REMARK
3
  L11:  5.4421 L22:  4.0606


REMARK
3
  L33:  1.5369 L12:  4.6706


REMARK
3
  L13: −2.0058 L23: −1.5537


REMARK
3
 S TENSOR


REMARK
3
  S11:  .1622 S12:  .0431 S13:  .3257


REMARK
3
  S21:  .1755 S22:  .0292 S23:  .2977


REMARK
3
  S31:  −.1910 S32:  −.0506 S33: −.1914


REMARK
3


REMARK
3
 TLS GROUP:  7


REMARK
3
 NUMBER OF COMPONENTS GROUP: 1


REMARK
3
 COMPONENTS  C SSSEQI TO C SSSEQI


REMARK
3
 RESIDUE RANGE: B  288  B  374


REMARK
3
 ORIGIN FOR THE GROUP (A): −41.6930 −10.8250 −19.6636


REMARK
3
 T TENSOR


REMARK
3
  T11:  .1424 T22:  .0604


REMARK
3
  T33:  .1153 T12:  .0184


REMARK
3
  T13:  .0146 T23:  .0276


REMARK
3
 L TENSOR


REMARK
3
  L11:  .6426 L22:  .8163


REMARK
3
  L33:  2.3437 L12:  −.1831


REMARK
3
  L13:  −.5246 L23:  .4917


REMARK
3
 S TENSOR


REMARK
3
  S11:  .0592 S12:  −.0206 S13:  .0071


REMARK
3
  S21:  .0906 S22:  −.0229 S23: −.1585


REMARK
3
  S31:  −.0355 S32:  .0262 S33: −.0363


REMARK
3


REMARK
3
 TLS GROUP:  8


REMARK
3
 NUMBER OF COMPONENTS GROUP: 1


REMARK
3
 OMPONENTS  C SSSEQI TO C SSSEQI


REMARK
3
 RESIDUE RANGE: B  375  B  541


REMARK
3
 ORIGIN FOR THE GROUP (A): −53.4886 −28.3212 −26.0670


REMARK
3
 T TENSOR


REMARK
3
  T11:  .1107 T22:  .1220


REMARK
3
  T33:  .1514 T12:  .0692


REMARK
3
  T13:  −.0073 T23:  .0518


REMARK
3
 L TENSOR


REMARK
3
  L11:  2.6766 L22:  1.8433


REMARK
3
  L33:  2.6389 L12:  .1130


REMARK
3
  L13: −2.4696 L23:  .6986


REMARK
3
 S TENSOR


REMARK
3
  S11:  .1115 S12:  .3882 S13:  .0569


REMARK
3
  S21:  −.0725 S22:  −.0724 S23:  .1450


REMARK
3
  S31:  −.1453 S32:  −.4044 S33: −.0392


REMARK
3


REMARK
3


REMARK
3
BULK SOLVENT MODELLING.


REMARK
3
 METHOD USED: MASK


REMARK
3
 PARAMETERS FOR MASK CALCULATION


REMARK
3
 VDW PROBE RADIUS : 1.40


REMARK
3
 ION PROBE RADIUS : .80


REMARK
3
 SHRINKAGE RADIUS : .80


REMARK
3


REMARK
3
OTHER REFINEMENT REMARKS:


REMARK
3
HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS


REMARK
3
U VALUES : RESIDUAL ONLY


REMARK
3


CISPEP
1
ALA A 446 ARG A 447    .00


CISPEP
2
GLY A 515 ASP A 516    .00


CISPEP
3
THR B 22 ASP B 23    .00


CISPEP
4
ALA B 446 ARG B 447    .00


CISPEP
5
GLY B 515 ASP B 516    .00








CRYST1
155.800 155.800 143.690 90.00 90.00 90.00 P 43 21 2












SCALE1
.006418
.000000
.000000
.00000



SCALE2
.000000
.006418
.000000
.00000


SCALE3
.000000
.000000
.006959
.00000


















ATOM
1
N
LEU
A
17
−63.930
24.416
−19.202
1.00
30.90
N


ATOM
2
CA
LEU
A
17
−64.132
23.019
−19.731
1.00
31.43
C


ATOM
4
CB
LEU
A
17
−63.308
22.800
−21.021
1.00
31.35
C


ATOM
7
CG
LEU
A
17
−64.002
23.016
−22.386
1.00
31.95
C


ATOM
9
CD1
LEU
A
17
−62.989
23.068
−23.550
1.00
31.88
C


ATOM
13
CD2
LEU
A
17
−65.052
21.915
−22.676
1.00
31.95
C


ATOM
17
C
LEU
A
17
−63.783
21.952
−18.660
1.00
31.39
C


ATOM
18
O
LEU
A
17
−63.142
22.291
−17.651
1.00
31.61
O


ATOM
22
N
LEU
A
18
−64.216
20.693
−18.877
1.00
31.11
N


ATOM
23
CA
LEU
A
18
−63.986
19.529
−17.957
1.00
30.78
C


ATOM
25
CB
LEU
A
18
−62.503
19.072
−17.935
1.00
30.97
C


ATOM
28
CG
LEU
A
18
−61.303
19.949
−17.496
1.00
31.10
C


ATOM
30
CD1
LEU
A
18
−61.406
20.430
−16.064
1.00
30.71
C


ATOM
34
CD2
LEU
A
18
−59.977
19.173
−17.703
1.00
31.87
C


ATOM
38
C
LEU
A
18
−64.531
19.665
−16.522
1.00
30.38
C


ATOM
39
O
LEU
A
18
−64.837
18.667
−15.883
1.00
29.86
O


ATOM
41
N
SER
A
19
−64.587
20.900
−16.023
1.00
30.45
N


ATOM
42
CA
SER
A
19
−65.335
21.276
−14.815
1.00
30.37
C


ATOM
44
CB
SER
A
19
−64.604
22.417
−14.026
1.00
30.02
C


ATOM
47
OG
SER
A
19
−64.881
23.741
−14.489
1.00
28.00
O


ATOM
49
C
SER
A
19
−66.784
21.654
−15.218
1.00
31.03
C


ATOM
50
O
SER
A
19
−67.666
21.711
−14.367
1.00
30.95
O


ATOM
52
N
SER
A
20
−67.023
21.880
−16.519
1.00
31.85
N


ATOM
53
CA
SER
A
20
−68.355
22.247
−17.051
1.00
32.37
C


ATOM
55
CB
SER
A
20
−68.291
22.507
−18.579
1.00
32.39
C


ATOM
58
OG
SER
A
20
−67.387
23.542
−18.931
1.00
31.77
O


ATOM
60
C
SER
A
20
−69.357
21.124
−16.744
1.00
33.04
C


ATOM
61
O
SER
A
20
−69.076
20.254
−15.922
1.00
33.08
O


ATOM
63
N
ASP
A
21
−70.522
21.136
−17.393
1.00
33.88
N


ATOM
64
CA
ASP
A
21
−71.512
20.069
−17.206
1.00
34.48
C


ATOM
66
CB
ASP
A
21
−72.907
20.655
−17.127
1.00
34.59
C


ATOM
69
CG
ASP
A
21
−73.022
21.661
−16.006
1.00
35.54
C


ATOM
70
OD1
ASP
A
21
−72.041
21.836
−15.251
1.00
35.31
O


ATOM
71
OD2
ASP
A
21
−74.082
22.289
−15.870
1.00
38.72
O


ATOM
72
C
ASP
A
21
−71.409
18.975
−18.260
1.00
34.94
C


ATOM
73
O
ASP
A
21
−72.134
18.947
−19.269
1.00
34.58
O


ATOM
75
N
THR
A
22
−70.457
18.086
−17.989
1.00
35.66
N


ATOM
76
CA
THR
A
22
−70.340
16.799
−18.657
1.00
36.27
C


ATOM
78
CB
THR
A
22
−68.895
16.576
−19.168
1.00
36.30
C


ATOM
80
OG1
THR
A
22
−67.968
17.278
−18.322
1.00
36.04
O


ATOM
82
CG2
THR
A
22
−68.755
17.088
−20.604
1.00
36.11
C


ATOM
86
C
THR
A
22
−70.792
15.714
−17.648
1.00
36.83
C


ATOM
87
O
THR
A
22
−69.968
15.022
−17.035
1.00
37.05
O


ATOM
89
N
ASP
A
23
−72.121
15.599
−17.494
1.00
37.27
N


ATOM
90
CA
ASP
A
23
−72.790
14.802
−16.441
1.00
37.25
C


ATOM
92
CB
ASP
A
23
−72.962
15.659
−15.167
1.00
37.17
C


ATOM
95
CG
ASP
A
23
−71.625
16.081
−14.549
1.00
37.74
C


ATOM
96
OD1
ASP
A
23
−70.714
15.241
−14.436
1.00
38.88
O


ATOM
97
OD2
ASP
A
23
−71.472
17.256
−14.164
1.00
38.91
O


ATOM
98
C
ASP
A
23
−74.172
14.351
−16.956
1.00
37.18
C


ATOM
99
O
ASP
A
23
−75.137
15.112
−16.846
1.00
37.10
O


ATOM
101
N
GLU
A
24
−74.280
13.134
−17.501
1.00
37.23
N


ATOM
102
CA
GLU
A
24
−75.406
12.815
−18.410
1.00
37.58
C


ATOM
104
CB
GLU
A
24
−74.941
13.007
−19.865
1.00
37.77
C


ATOM
107
CG
GLU
A
24
−74.424
14.417
−20.239
1.00
37.98
C


ATOM
110
CD
GLU
A
24
−74.121
14.556
−21.746
1.00
38.35
C


ATOM
111
OE1
GLU
A
24
−74.284
13.564
−22.503
1.00
38.12
O


ATOM
112
OE2
GLU
A
24
−73.721
15.661
−22.174
1.00
38.18
O


ATOM
113
C
GLU
A
24
−76.139
11.440
−18.323
1.00
37.79
C


ATOM
114
O
GLU
A
24
−77.323
11.397
−17.960
1.00
37.90
O


ATOM
116
N
SER
A
25
−75.462
10.345
−18.696
1.00
37.92
N


ATOM
117
CA
SER
A
25
−76.140
9.070
−19.048
1.00
38.09
C


ATOM
119
CB
SER
A
25
−75.167
8.076
−19.723
1.00
38.11
C


ATOM
122
OG
SER
A
25
−74.503
7.239
−18.787
1.00
38.02
O


ATOM
124
C
SER
A
25
−76.917
8.374
−17.907
1.00
38.42
C


ATOM
125
O
SER
A
25
−76.635
8.581
−16.724
1.00
38.15
O


ATOM
127
N
ILE
A
26
−77.861
7.511
−18.312
1.00
39.00
N


ATOM
128
CA
ILE
A
26
−78.993
7.042
−17.476
1.00
39.30
C


ATOM
130
CB
ILE
A
26
−78.594
5.965
−16.429
1.00
39.34
C


ATOM
132
CG1
ILE
A
26
−77.892
4.791
−17.120
1.00
39.37
C


ATOM
135
CD1
ILE
A
26
−77.766
3.531
−16.253
1.00
39.46
C


ATOM
139
CG2
ILE
A
26
−79.836
5.423
−15.714
1.00
39.29
C


ATOM
143
C
ILE
A
26
−79.716
8.260
−16.852
1.00
39.66
C


ATOM
144
O
ILE
A
26
−79.274
8.831
−15.838
1.00
39.49
O


ATOM
146
N
GLU
A
27
−80.837
8.623
−17.486
1.00
39.99
N


ATOM
147
CA
GLU
A
27
−81.450
9.965
−17.383
1.00
40.11
C


ATOM
149
CB
GLU
A
27
−82.395
10.183
−18.587
1.00
40.19
C


ATOM
152
CG
GLU
A
27
−81.632
10.311
−19.904
1.00
40.60
C


ATOM
155
CD
GLU
A
27
−82.528
10.434
−21.116
1.00
41.06
C


ATOM
156
OE1
GLU
A
27
−83.328
9.502
−21.367
1.00
41.10
O


ATOM
157
OE2
GLU
A
27
−82.409
11.458
−21.830
1.00
41.36
O


ATOM
158
C
GLU
A
27
−82.144
10.264
−16.042
1.00
40.02
C


ATOM
159
O
GLU
A
27
−81.977
9.518
−15.071
1.00
40.01
O


ATOM
161
N
VAL
A
28
−82.899
11.370
−16.005
1.00
39.84
N


ATOM
162
CA
VAL
A
28
−83.463
11.960
−14.779
1.00
39.65
C


ATOM
164
CB
VAL
A
28
−83.884
10.905
−13.694
1.00
39.78
C


ATOM
166
CG1
VAL
A
28
−84.472
11.596
−12.456
1.00
39.87
C


ATOM
170
CG2
VAL
A
28
−84.887
9.880
−14.274
1.00
39.49
C


ATOM
174
C
VAL
A
28
−82.469
12.980
−14.206
1.00
39.46
C


ATOM
175
O
VAL
A
28
−82.798
13.717
−13.270
1.00
39.38
O


ATOM
177
N
HIS
A
29
−81.264
13.025
−14.786
1.00
39.30
N


ATOM
178
CA
HIS
A
29
−80.236
14.006
−14.412
1.00
39.23
C


ATOM
180
CB
HIS
A
29
−78.866
13.344
−14.186
1.00
39.45
C


ATOM
183
CG
HIS
A
29
−78.910
12.139
−13.296
1.00
41.29
C


ATOM
184
ND1
HIS
A
29
−78.407
12.140
−12.007
1.00
42.51
N


ATOM
186
CE1
HIS
A
29
−78.585
10.942
−11.472
1.00
43.37
C


ATOM
188
NE2
HIS
A
29
−79.181
10.164
−12.365
1.00
43.18
N


ATOM
190
CD2
HIS
A
29
−79.393
10.888
−13.515
1.00
42.55
C


ATOM
192
C
HIS
A
29
−80.090
15.110
−15.464
1.00
38.60
C


ATOM
193
O
HIS
A
29
−79.049
15.774
−15.499
1.00
38.61
O


ATOM
195
N
LYS
A
30
−81.110
15.320
−16.311
1.00
37.87
N


ATOM
196
CA
LYS
A
30
−81.265
16.610
−17.001
1.00
37.22
C


ATOM
198
CB
LYS
A
30
−82.283
16.542
−18.140
1.00
37.09
C


ATOM
201
CG
LYS
A
30
−81.764
15.794
−19.360
1.00
37.00
C


ATOM
204
CD
LYS
A
30
−82.831
15.653
−20.457
1.00
36.89
C


ATOM
207
CE
LYS
A
30
−82.413
14.637
−21.531
1.00
36.48
C


ATOM
210
NZ
LYS
A
30
−83.422
14.458
−22.611
1.00
35.46
N


ATOM
214
C
LYS
A
30
−81.648
17.649
−15.938
1.00
36.88
C


ATOM
215
O
LYS
A
30
−82.516
18.499
−16.136
1.00
36.71
O


ATOM
217
N
ASP
A
31
−80.982
17.506
−14.788
1.00
36.58
N


ATOM
218
CA
ASP
A
31
−80.895
18.476
−13.720
1.00
36.18
C


ATOM
220
CB
ASP
A
31
−80.518
17.751
−12.396
1.00
36.20
C


ATOM
223
CG
ASP
A
31
−79.863
18.669
−11.342
1.00
36.65
C


ATOM
224
OD1
ASP
A
31
−80.055
19.901
−11.390
1.00
38.17
O


ATOM
225
OD2
ASP
A
31
−79.147
18.150
−10.449
1.00
35.46
O


ATOM
226
C
ASP
A
31
−79.811
19.426
−14.226
1.00
35.74
C


ATOM
227
O
ASP
A
31
−78.628
19.279
−13.918
1.00
35.44
O


ATOM
229
N
LYS
A
32
−80.220
20.343
−15.096
1.00
35.30
N


ATOM
230
CA
LYS
A
32
−79.360
21.449
−15.507
1.00
34.69
C


ATOM
232
CB
LYS
A
32
−78.698
21.188
−16.849
1.00
34.63
C


ATOM
235
CG
LYS
A
32
−77.699
20.042
−16.765
1.00
34.51
C


ATOM
238
CD
LYS
A
32
−76.953
19.853
−18.078
1.00
34.48
C


ATOM
241
CE
LYS
A
32
−76.859
18.387
−18.503
1.00
33.77
C


ATOM
244
NZ
LYS
A
32
−77.002
18.244
−19.985
1.00
32.92
N


ATOM
248
C
LYS
A
32
−80.162
22.745
−15.459
1.00
34.01
C


ATOM
249
O
LYS
A
32
−80.468
23.380
−16.473
1.00
33.41
O


ATOM
251
N
ALA
A
33
−80.540
23.051
−14.219
1.00
33.21
N


ATOM
252
CA
ALA
A
33
−80.785
24.381
−13.766
1.00
32.60
C


ATOM
254
CB
ALA
A
33
−81.619
24.347
−12.484
1.00
32.28
C


ATOM
258
C
ALA
A
33
−79.411
25.031
−13.524
1.00
32.31
C


ATOM
259
O
ALA
A
33
−79.335
26.081
−12.901
1.00
32.80
O


ATOM
261
N
LYS
A
34
−78.323
24.389
−13.968
1.00
31.70
N


ATOM
262
CA
LYS
A
34
−77.035
25.071
−14.186
1.00
31.12
C


ATOM
264
CB
LYS
A
34
−75.863
24.089
−14.128
1.00
31.23
C


ATOM
267
CG
LYS
A
34
−75.118
24.077
−12.791
1.00
31.76
C


ATOM
270
CD
LYS
A
34
−73.584
24.412
−12.921
1.00
31.49
C


ATOM
273
CE
LYS
A
34
−72.685
23.269
−12.506
1.00
30.50
C


ATOM
276
NZ
LYS
A
34
−72.951
22.090
−13.348
1.00
29.53
N


ATOM
280
C
LYS
A
34
−77.024
25.884
−15.518
1.00
30.50
C


ATOM
281
O
LYS
A
34
−76.977
25.353
−16.625
1.00
29.82
O


ATOM
283
N
LYS
A
35
−76.927
27.191
−15.337
1.00
29.97
N


ATOM
284
CA
LYS
A
35
−77.632
28.233
−16.098
1.00
29.41
C


ATOM
286
CB
LYS
A
35
−78.852
27.704
−16.867
1.00
29.53
C


ATOM
289
CG
LYS
A
35
−80.166
27.594
−16.077
1.00
30.18
C


ATOM
292
CD
LYS
A
35
−81.030
28.858
−16.203
1.00
31.25
C


ATOM
295
CE
LYS
A
35
−82.375
28.721
−15.486
1.00
31.85
C


ATOM
298
NZ
LYS
A
35
−83.298
29.865
−15.783
1.00
31.73
N


ATOM
302
C
LYS
A
35
−78.053
29.277
−15.026
1.00
28.53
C


ATOM
303
O
LYS
A
35
−78.246
30.460
−15.307
1.00
28.53
O


ATOM
305
N
LEU
A
36
−78.225
28.791
−13.796
1.00
27.29
N


ATOM
306
CA
LEU
A
36
−77.998
29.576
−12.594
1.00
26.21
C


ATOM
308
CB
LEU
A
36
−78.088
28.679
−11.364
1.00
25.76
C


ATOM
311
CG
LEU
A
36
−79.468
28.350
−10.832
1.00
24.08
C


ATOM
313
CD1
LEU
A
36
−79.376
27.233
−9.836
1.00
22.16
C


ATOM
317
CD2
LEU
A
36
−80.051
29.586
−10.213
1.00
23.45
C


ATOM
321
C
LEU
A
36
−76.587
30.137
−12.665
1.00
25.93
C


ATOM
322
O
LEU
A
36
−76.290
31.221
−12.137
1.00
26.11
O


ATOM
324
N
GLU
A
37
−75.714
29.332
−13.260
1.00
25.24
N


ATOM
325
CA
GLU
A
37
−74.381
29.739
−13.650
1.00
24.85
C


ATOM
327
CB
GLU
A
37
−73.678
28.566
−14.313
1.00
24.74
C


ATOM
330
CG
GLU
A
37
−72.182
28.667
−14.285
1.00
23.93
C


ATOM
333
CD
GLU
A
37
−71.519
27.490
−14.937
1.00
22.32
C


ATOM
334
OE1
GLU
A
37
−72.203
26.738
−15.657
1.00
20.57
O


ATOM
335
OE2
GLU
A
37
−70.303
27.322
−14.728
1.00
22.27
O


ATOM
336
C
GLU
A
37
−74.391
30.927
−14.600
1.00
24.69
C


ATOM
337
O
GLU
A
37
−73.666
31.891
−14.389
1.00
24.49
O


ATOM
339
N
ALA
A
38
−75.203
30.850
−15.650
1.00
24.68
N


ATOM
340
CA
ALA
A
38
−75.352
31.960
−16.599
1.00
24.87
C


ATOM
342
CB
ALA
A
38
−76.453
31.656
−17.609
1.00
24.53
C


ATOM
346
C
ALA
A
38
−75.643
33.278
−15.880
1.00
25.08
C


ATOM
347
O
ALA
A
38
−75.009
34.295
−16.135
1.00
25.21
O


ATOM
349
N
GLU
A
39
−76.591
33.228
−14.956
1.00
25.35
N


ATOM
350
CA
GLU
A
39
−77.050
34.400
−14.219
1.00
25.51
C


ATOM
352
CB
GLU
A
39
−78.283
34.014
−13.396
1.00
25.95
C


ATOM
355
CG
GLU
A
39
−79.302
35.119
−13.136
1.00
27.03
C


ATOM
358
CD
GLU
A
39
−80.715
34.557
−12.906
1.00
28.70
C


ATOM
359
OE1
GLU
A
39
−81.006
33.416
−13.346
1.00
27.40
O


ATOM
360
OE2
GLU
A
39
−81.540
35.266
−12.289
1.00
31.12
O


ATOM
361
C
GLU
A
39
−75.970
34.966
−13.306
1.00
25.11
C


ATOM
362
O
GLU
A
39
−75.870
36.167
−13.164
1.00
25.28
O


ATOM
364
N
VAL
A
40
−75.182
34.107
−12.672
1.00
24.86
N


ATOM
365
CA
VAL
A
40
−74.079
34.568
−11.824
1.00
24.69
C


ATOM
367
CB
VAL
A
40
−73.511
33.420
−10.952
1.00
24.56
C


ATOM
369
CG1
VAL
A
40
−72.239
33.857
−10.240
1.00
23.69
C


ATOM
373
CG2
VAL
A
40
−74.553
32.955
−9.955
1.00
24.68
C


ATOM
377
C
VAL
A
40
−72.971
35.148
−12.698
1.00
24.76
C


ATOM
378
O
VAL
A
40
−72.337
36.139
−12.362
1.00
24.16
O


ATOM
380
N
ARG
A
41
−72.744
34.506
−13.831
1.00
25.25
N


ATOM
381
CA
ARG
A
41
−71.727
34.948
−14.769
1.00
25.66
C


ATOM
383
CB
ARG
A
41
−71.576
33.927
−15.896
1.00
25.88
C


ATOM
386
CG
ARG
A
41
−70.726
34.385
−17.062
1.00
27.04
C


ATOM
389
CD
ARG
A
41
−71.519
35.178
−18.095
1.00
27.90
C


ATOM
392
NE
ARG
A
41
−70.653
35.635
−19.180
1.00
29.20
N


ATOM
394
CZ
ARG
A
41
−70.946
36.615
−20.034
1.00
29.94
C


ATOM
395
NH1
ARG
A
41
−70.077
36.945
−20.980
1.00
29.86
N


ATOM
398
NH2
ARG
A
41
−72.096
37.273
−19.957
1.00
30.54
N


ATOM
401
C
ARG
A
41
−72.104
36.301
−15.335
1.00
25.60
C


ATOM
402
O
ARG
A
41
−71.237
37.113
−15.612
1.00
25.77
O


ATOM
404
N
ARG
A
42
−73.400
36.521
−15.537
1.00
25.50
N


ATOM
405
CA
ARG
A
42
−73.900
37.810
−15.979
1.00
25.39
C


ATOM
407
CB
ARG
A
42
−75.389
37.723
−16.313
1.00
25.09
C


ATOM
410
CG
ARG
A
42
−76.077
39.062
−16.445
1.00
23.84
C


ATOM
413
CD
ARG
A
42
−77.459
38.906
−16.980
1.00
22.37
C


ATOM
416
NE
ARG
A
42
−78.448
38.554
−15.965
1.00
21.69
N


ATOM
418
CZ
ARG
A
42
−79.705
38.206
−16.248
1.00
22.99
C


ATOM
419
NH1
ARG
A
42
−80.122
38.146
−17.517
1.00
24.16
N


ATOM
422
NH2
ARG
A
42
−80.557
37.900
−15.275
1.00
22.94
N


ATOM
425
C
ARG
A
42
−73.666
38.889
−14.925
1.00
26.00
C


ATOM
426
O
ARG
A
42
−73.173
39.959
−15.244
1.00
26.17
O


ATOM
428
N
GLU
A
43
−74.014
38.621
−13.673
1.00
26.66
N


ATOM
429
CA
GLU
A
43
−73.930
39.662
−12.650
1.00
27.47
C


ATOM
431
CB
GLU
A
43
−74.749
39.304
−11.401
1.00
27.97
C


ATOM
434
CG
GLU
A
43
−76.274
39.266
−11.661
1.00
31.05
C


ATOM
437
CD
GLU
A
43
−76.869
40.636
−12.063
1.00
34.68
C


ATOM
438
OE1
GLU
A
43
−76.804
41.571
−11.222
1.00
37.63
O


ATOM
439
OE2
GLU
A
43
−77.398
40.773
−13.205
1.00
34.52
O


ATOM
440
C
GLU
A
43
−72.494
40.035
−12.275
1.00
27.15
C


ATOM
441
O
GLU
A
43
−72.292
41.118
−11.716
1.00
27.49
O


ATOM
443
N
ILE
A
44
−71.517
39.166
−12.588
1.00
26.57
N


ATOM
444
CA
ILE
A
44
−70.090
39.456
−12.334
1.00
25.80
C


ATOM
446
CB
ILE
A
44
−69.206
38.187
−12.187
1.00
25.52
C


ATOM
448
CG1
ILE
A
44
−69.624
37.320
−11.010
1.00
24.69
C


ATOM
451
CD1
ILE
A
44
−68.828
36.061
−10.899
1.00
23.08
C


ATOM
455
CG2
ILE
A
44
−67.790
38.581
−11.916
1.00
25.73
C


ATOM
459
C
ILE
A
44
−69.522
40.286
−13.472
1.00
25.40
C


ATOM
460
O
ILE
A
44
−68.745
41.211
−13.236
1.00
25.26
O


ATOM
462
N
ASN
A
45
−69.912
39.945
−14.700
1.00
25.14
N


ATOM
463
CA
ASN
A
45
−69.480
40.669
−15.906
1.00
25.02
C


ATOM
465
CB
ASN
A
45
−69.696
39.813
−17.151
1.00
24.69
C


ATOM
468
CG
ASN
A
45
−68.662
38.735
−17.302
1.00
24.01
C


ATOM
469
OD1
ASN
A
45
−67.470
39.013
−17.395
1.00
23.81
O


ATOM
470
ND2
ASN
A
45
−69.111
37.490
−17.344
1.00
23.28
N


ATOM
473
C
ASN
A
45
−70.177
42.014
−16.114
1.00
25.52
C


ATOM
474
O
ASN
A
45
−69.704
42.836
−16.890
1.00
25.40
O


ATOM
476
N
ASN
A
46
−71.308
42.210
−15.437
1.00
26.33
N


ATOM
477
CA
ASN
A
46
−72.085
43.453
−15.468
1.00
26.72
C


ATOM
479
CB
ASN
A
46
−73.102
43.404
−14.322
1.00
26.31
C


ATOM
482
CG
ASN
A
46
−73.935
44.641
−14.209
1.00
24.24
C


ATOM
483
OD1
ASN
A
46
−73.948
45.472
−15.091
1.00
22.38
O


ATOM
484
ND2
ASN
A
46
−74.645
44.766
−13.106
1.00
21.64
N


ATOM
487
C
ASN
A
46
−71.185
44.680
−15.349
1.00
28.11
C


ATOM
488
O
ASN
A
46
−70.603
44.933
−14.304
1.00
28.16
O


ATOM
490
N
GLU
A
47
−71.084
45.441
−16.433
1.00
29.93
N


ATOM
491
CA
GLU
A
47
−70.128
46.558
−16.544
1.00
31.30
C


ATOM
493
CB
GLU
A
47
−69.933
46.969
−18.013
1.00
31.28
C


ATOM
496
CG
GLU
A
47
−69.737
45.795
−19.020
1.00
32.37
C


ATOM
499
CD
GLU
A
47
−71.060
45.127
−19.545
1.00
32.69
C


ATOM
500
OE1
GLU
A
47
−72.055
45.833
−19.816
1.00
32.16
O


ATOM
501
OE2
GLU
A
47
−71.087
43.880
−19.699
1.00
32.41
O


ATOM
502
C
GLU
A
47
−70.582
47.774
−15.729
1.00
32.47
C


ATOM
503
O
GLU
A
47
−69.760
48.525
−15.230
1.00
32.86
O


ATOM
505
N
LYS
A
48
−71.898
47.945
−15.585
1.00
33.93
N


ATOM
506
CA
LYS
A
48
−72.491
49.075
−14.861
1.00
34.85
C


ATOM
508
CB
LYS
A
48
−73.660
49.672
−15.684
1.00
35.08
C


ATOM
511
CG
LYS
A
48
−73.221
50.119
−17.126
1.00
36.58
C


ATOM
514
CD
LYS
A
48
−74.035
51.277
−17.751
1.00
37.93
C


ATOM
517
CE
LYS
A
48
−75.353
50.799
−18.415
1.00
39.02
C


ATOM
520
NZ
LYS
A
48
−75.159
49.990
−19.669
1.00
39.11
N


ATOM
524
C
LYS
A
48
−72.926
48.632
−13.465
1.00
35.26
C


ATOM
525
O
LYS
A
48
−74.011
48.954
−13.015
1.00
35.04
O


ATOM
527
N
ALA
A
49
−72.051
47.890
−12.790
1.00
36.29
N


ATOM
528
CA
ALA
A
49
−72.321
47.348
−11.456
1.00
37.18
C


ATOM
530
CB
ALA
A
49
−71.828
45.923
−11.353
1.00
37.20
C


ATOM
534
C
ALA
A
49
−71.618
48.191
−10.418
1.00
37.88
C


ATOM
535
O
ALA
A
49
−70.480
48.610
−10.638
1.00
38.17
O


ATOM
537
N
GLU
A
50
−72.278
48.404
−9.280
1.00
38.53
N


ATOM
538
CA
GLU
A
50
−71.730
49.242
−8.213
1.00
39.08
C


ATOM
540
CB
GLU
A
50
−72.790
49.545
−7.146
1.00
39.43
C


ATOM
543
CG
GLU
A
50
−72.708
50.953
−6.600
1.00
40.86
C


ATOM
546
CD
GLU
A
50
−73.089
52.010
−7.639
1.00
42.77
C


ATOM
547
OE1
GLU
A
50
−74.225
52.530
−7.570
1.00
45.07
O


ATOM
548
OE2
GLU
A
50
−72.265
52.320
−8.530
1.00
43.35
O


ATOM
549
C
GLU
A
50
−70.560
48.521
−7.600
1.00
38.89
C


ATOM
550
O
GLU
A
50
−70.699
47.378
−7.201
1.00
39.05
O


ATOM
552
N
PHE
A
51
−69.411
49.181
−7.536
1.00
38.99
N


ATOM
553
CA
PHE
A
51
−68.161
48.502
−7.196
1.00
39.52
C


ATOM
555
CB
PHE
A
51
−67.000
49.494
−7.098
1.00
40.05
C


ATOM
558
CG
PHE
A
51
−66.460
49.948
−8.431
1.00
42.80
C


ATOM
559
CD1
PHE
A
51
−66.135
49.014
−9.435
1.00
45.21
C


ATOM
561
CE1
PHE
A
51
−65.612
49.425
−10.675
1.00
46.12
C


ATOM
563
CZ
PHE
A
51
−65.403
50.790
−10.917
1.00
47.21
C


ATOM
565
CE2
PHE
A
51
−65.726
51.741
−9.913
1.00
46.72
C


ATOM
567
CD2
PHE
A
51
−66.247
51.310
−8.679
1.00
45.05
C


ATOM
569
C
PHE
A
51
−68.223
47.687
−5.905
1.00
39.05
C


ATOM
570
O
PHE
A
51
−67.897
46.506
−5.900
1.00
39.28
O


ATOM
572
N
LEU
A
52
−68.648
48.307
−4.813
1.00
38.42
N


ATOM
573
CA
LEU
A
52
−68.709
47.610
−3.528
1.00
37.94
C


ATOM
575
CB
LEU
A
52
−69.084
48.594
−2.413
1.00
38.53
C


ATOM
578
CG
LEU
A
52
−68.057
49.726
−2.181
1.00
40.46
C


ATOM
580
CD1
LEU
A
52
−68.691
51.101
−1.787
1.00
41.88
C


ATOM
584
CD2
LEU
A
52
−67.020
49.274
−1.141
1.00
41.83
C


ATOM
588
C
LEU
A
52
−69.667
46.406
−3.532
1.00
36.72
C


ATOM
589
O
LEU
A
52
−69.517
45.494
−2.724
1.00
36.84
O


ATOM
591
N
THR
A
53
−70.649
46.398
−4.431
1.00
35.37
N


ATOM
592
CA
THR
A
53
−71.545
45.247
−4.569
1.00
34.11
C


ATOM
594
CB
THR
A
53
−72.867
45.594
−5.277
1.00
33.87
C


ATOM
596
OG1
THR
A
53
−73.339
46.863
−4.828
1.00
33.49
O


ATOM
598
CG2
THR
A
53
−73.917
44.535
−4.983
1.00
33.70
C


ATOM
602
C
THR
A
53
−70.853
44.148
−5.359
1.00
33.04
C


ATOM
603
O
THR
A
53
−70.893
42.977
−4.973
1.00
32.90
O


ATOM
605
N
LEU
A
54
−70.228
44.528
−6.467
1.00
31.76
N


ATOM
606
CA
LEU
A
54
−69.469
43.578
−7.286
1.00
31.13
C


ATOM
608
CB
LEU
A
54
−68.721
44.308
−8.392
1.00
30.92
C


ATOM
611
CG
LEU
A
54
−68.028
43.437
−9.424
1.00
30.27
C


ATOM
613
CD1
LEU
A
54
−69.034
42.769
−10.310
1.00
29.56
C


ATOM
617
CD2
LEU
A
54
−67.108
44.306
−10.242
1.00
31.15
C


ATOM
621
C
LEU
A
54
−68.466
42.808
−6.443
1.00
30.60
C


ATOM
622
O
LEU
A
54
−68.378
41.587
−6.543
1.00
30.30
O


ATOM
624
N
LEU
A
55
−67.725
43.551
−5.618
1.00
30.09
N


ATOM
625
CA
LEU
A
55
−66.730
43.000
−4.695
1.00
29.47
C


ATOM
627
CB
LEU
A
55
−66.006
44.126
−3.944
1.00
29.44
C


ATOM
630
CG
LEU
A
55
−65.069
45.017
−4.781
1.00
29.74
C


ATOM
632
CD1
LEU
A
55
−64.609
46.255
−4.003
1.00
29.43
C


ATOM
636
CD2
LEU
A
55
−63.859
44.234
−5.286
1.00
29.64
C


ATOM
640
C
LEU
A
55
−67.340
42.024
−3.696
1.00
28.87
C


ATOM
641
O
LEU
A
55
−66.746
40.991
−3.383
1.00
28.98
O


ATOM
643
N
GLU
A
56
−68.525
42.330
−3.198
1.00
28.01
N


ATOM
644
CA
GLU
A
56
−69.160
41.419
−2.256
1.00
27.64
C


ATOM
646
CB
GLU
A
56
−70.172
42.173
−1.395
1.00
28.19
C


ATOM
649
CG
GLU
A
56
−69.497
43.167
−.459
1.00
29.75
C


ATOM
652
CD
GLU
A
56
−70.457
43.858
.478
1.00
33.04
C


ATOM
653
OE1
GLU
A
56
−71.667
43.538
.481
1.00
34.46
O


ATOM
654
OE2
GLU
A
56
−69.988
44.737
1.226
1.00
36.78
O


ATOM
655
C
GLU
A
56
−69.776
40.200
−2.945
1.00
26.37
C


ATOM
656
O
GLU
A
56
−69.914
39.146
−2.333
1.00
26.02
O


ATOM
658
N
LEU
A
57
−70.134
40.347
−4.218
1.00
25.30
N


ATOM
659
CA
LEU
A
57
−70.569
39.212
−5.036
1.00
24.17
C


ATOM
661
CB
LEU
A
57
−71.125
39.680
−6.382
1.00
23.71
C


ATOM
664
CG
LEU
A
57
−71.417
38.568
−7.390
1.00
22.27
C


ATOM
666
CD1
LEU
A
57
−72.515
37.675
−6.858
1.00
19.31
C


ATOM
670
CD2
LEU
A
57
−71.752
39.167
−8.768
1.00
20.30
C


ATOM
674
C
LEU
A
57
−69.404
38.267
−5.284
1.00
23.69
C


ATOM
675
O
LEU
A
57
−69.547
37.051
−5.136
1.00
23.83
O


ATOM
677
N
ILE
A
58
−68.261
38.822
−5.682
1.00
22.82
N


ATOM
678
CA
ILE
A
58
−67.070
38.019
−5.925
1.00
22.39
C


ATOM
680
CB
ILE
A
58
−65.884
38.899
−6.355
1.00
22.21
C


ATOM
682
CG1
ILE
A
58
−66.088
39.386
−7.792
1.00
21.83
C


ATOM
685
CD1
ILE
A
58
−65.002
40.314
−8.320
1.00
20.43
C


ATOM
689
CG2
ILE
A
58
−64.565
38.131
−6.245
1.00
22.64
C


ATOM
693
C
ILE
A
58
−66.719
37.227
−4.662
1.00
22.35
C


ATOM
694
O
ILE
A
58
−66.520
36.002
−4.706
1.00
21.95
O


ATOM
696
N
ASP
A
59
−66.676
37.951
−3.543
1.00
22.32
N


ATOM
697
CA
ASP
A
59
−66.396
37.396
−2.215
1.00
22.26
C


ATOM
699
CB
ASP
A
59
−66.570
38.500
−1.171
1.00
22.67
C


ATOM
702
CG
ASP
A
59
−66.126
38.092
.213
1.00
23.91
C


ATOM
703
OD1
ASP
A
59
−65.271
37.182
.338
1.00
23.77
O


ATOM
704
OD2
ASP
A
59
−66.641
38.718
1.178
1.00
27.12
O


ATOM
705
C
ASP
A
59
−67.304
36.216
−1.901
1.00
21.79
C


ATOM
706
O
ASP
A
59
−66.828
35.102
−1.668
1.00
21.71
O


ATOM
708
N
ASN
A
60
−68.610
36.466
−1.926
1.00
21.51
N


ATOM
709
CA
ASN
A
60
−69.619
35.418
−1.759
1.00
21.28
C


ATOM
711
CB
ASN
A
60
−71.012
36.006
−1.958
1.00
21.25
C


ATOM
714
CG
ASN
A
60
−71.476
36.827
−.776
1.00
21.82
C


ATOM
715
OD1
ASN
A
60
−71.067
36.598
.364
1.00
20.81
O


ATOM
716
ND2
ASN
A
60
−72.361
37.783
−1.043
1.00
23.63
N


ATOM
719
C
ASN
A
60
−69.445
34.240
−2.725
1.00
21.22
C


ATOM
720
O
ASN
A
60
−69.444
33.053
−2.303
1.00
20.95
O


ATOM
722
N
VAL
A
61
−69.308
34.566
−4.018
1.00
20.83
N


ATOM
723
CA
VAL
A
61
−69.147
33.536
−5.043
1.00
20.57
C


ATOM
725
CB
VAL
A
61
−68.915
34.122
−6.466
1.00
20.53
C


ATOM
727
CG1
VAL
A
61
−68.382
33.042
−7.430
1.00
19.44
C


ATOM
731
CG2
VAL
A
61
−70.190
34.758
−7.006
1.00
19.75
C


ATOM
735
C
VAL
A
61
−67.981
32.646
−4.644
1.00
20.62
C


ATOM
736
O
VAL
A
61
−68.080
31.431
−4.699
1.00
20.56
O


ATOM
738
N
GLN
A
62
−66.888
33.255
−4.209
1.00
20.76
N


ATOM
739
CA
GLN
A
62
−65.703
32.481
−3.898
1.00
21.18
C


ATOM
741
CB
GLN
A
62
−64.461
33.379
−3.824
1.00
21.39
C


ATOM
744
CG
GLN
A
62
−64.007
33.920
−5.165
1.00
21.64
C


ATOM
747
CD
GLN
A
62
−62.608
34.512
−5.135
1.00
22.04
C


ATOM
748
OE1
GLN
A
62
−61.890
34.483
−6.145
1.00
23.18
O


ATOM
749
NE2
GLN
A
62
−62.217
35.062
−3.989
1.00
20.39
N


ATOM
752
C
GLN
A
62
−65.891
31.681
−2.607
1.00
21.09
C


ATOM
753
O
GLN
A
62
−65.544
30.485
−2.554
1.00
21.30
O


ATOM
755
N
ARG
A
63
−66.448
32.325
−1.583
1.00
20.63
N


ATOM
756
CA
ARG
A
63
−66.590
31.677
−.273
1.00
20.54
C


ATOM
758
CB
ARG
A
63
−67.025
32.686
.786
1.00
20.57
C


ATOM
761
CG
ARG
A
63
−66.031
33.823
.930
1.00
22.55
C


ATOM
764
CD
ARG
A
63
−66.214
34.632
2.179
1.00
24.93
C


ATOM
767
NE
ARG
A
63
−66.123
33.781
3.355
1.00
27.75
N


ATOM
769
CZ
ARG
A
63
−66.496
34.151
4.573
1.00
30.24
C


ATOM
770
NH1
ARG
A
63
−66.971
35.385
4.782
1.00
31.79
N


ATOM
773
NH2
ARG
A
63
−66.399
33.285
5.581
1.00
30.07
N


ATOM
776
C
ARG
A
63
−67.557
30.499
−.338
1.00
19.81
C


ATOM
777
O
ARG
A
63
−67.283
29.431
.209
1.00
19.67
O


ATOM
779
N
LEU
A
64
−68.661
30.684
−1.057
1.00
19.04
N


ATOM
780
CA
LEU
A
64
−69.636
29.617
−1.246
1.00
18.44
C


ATOM
782
CB
LEU
A
64
−70.865
30.167
−1.954
1.00
18.59
C


ATOM
785
CG
LEU
A
64
−71.662
31.170
−1.140
1.00
18.32
C


ATOM
787
CD1
LEU
A
64
−72.669
31.852
−2.034
1.00
18.10
C


ATOM
791
CD2
LEU
A
64
−72.334
30.444
.004
1.00
18.29
C


ATOM
795
C
LEU
A
64
−69.089
28.436
−2.037
1.00
17.80
C


ATOM
796
O
LEU
A
64
−69.802
27.453
−2.252
1.00
17.79
O


ATOM
798
N
GLY
A
65
−67.850
28.573
−2.515
1.00
17.07
N


ATOM
799
CA
GLY
A
65
−67.084
27.480
−3.081
1.00
16.27
C


ATOM
802
C
GLY
A
65
−67.137
27.406
−4.588
1.00
15.77
C


ATOM
803
O
GLY
A
65
−66.893
26.347
−5.154
1.00
15.78
O


ATOM
805
N
LEU
A
66
−67.439
28.521
−5.246
1.00
15.24
N


ATOM
806
CA
LEU
A
66
−67.621
28.537
−6.701
1.00
14.93
C


ATOM
808
CB
LEU
A
66
−68.977
29.146
−7.021
1.00
14.84
C


ATOM
811
CG
LEU
A
66
−70.204
28.334
−6.619
1.00
13.97
C


ATOM
813
CD1
LEU
A
66
−71.391
29.260
−6.650
1.00
13.74
C


ATOM
817
CD2
LEU
A
66
−70.415
27.125
−7.534
1.00
11.22
C


ATOM
821
C
LEU
A
66
−66.533
29.314
−7.454
1.00
15.09
C


ATOM
822
O
LEU
A
66
−66.621
29.497
−8.678
1.00
14.81
O


ATOM
824
N
GLY
A
67
−65.510
29.758
−6.720
1.00
15.21
N


ATOM
825
CA
GLY
A
67
−64.404
30.526
−7.280
1.00
14.98
C


ATOM
828
C
GLY
A
67
−63.873
29.952
−8.569
1.00
14.60
C


ATOM
829
O
GLY
A
67
−63.891
30.619
−9.579
1.00
14.63
O


ATOM
831
N
TYR
A
68
−63.419
28.706
−8.523
1.00
14.57
N


ATOM
832
CA
TYR
A
68
−62.831
28.029
−9.688
1.00
14.72
C


ATOM
834
CB
TYR
A
68
−62.608
26.539
−9.372
1.00
14.73
C


ATOM
837
CG
TYR
A
68
−63.858
25.689
−9.330
1.00
12.47
C


ATOM
838
CD1
TYR
A
68
−64.163
24.826
−10.366
1.00
10.68
C


ATOM
840
CE1
TYR
A
68
−65.310
24.043
−10.340
1.00
10.55
C


ATOM
842
CZ
TYR
A
68
−66.163
24.119
−9.258
1.00
10.41
C


ATOM
843
OH
TYR
A
68
−67.311
23.349
−9.212
1.00
6.97
O


ATOM
845
CE2
TYR
A
68
−65.864
24.979
−8.210
1.00
11.44
C


ATOM
847
CD2
TYR
A
68
−64.722
25.750
−8.252
1.00
11.29
C


ATOM
849
C
TYR
A
68
−63.648
28.141
−10.980
1.00
15.31
C


ATOM
850
O
TYR
A
68
−63.106
28.230
−12.083
1.00
15.01
O


ATOM
852
N
ARG
A
69
−64.959
28.136
−10.816
1.00
16.03
N


ATOM
853
CA
ARG
A
69
−65.884
28.083
−11.922
1.00
16.67
C


ATOM
855
CB
ARG
A
69
−67.224
27.649
−11.348
1.00
16.58
C


ATOM
858
CG
ARG
A
69
−68.332
27.445
−12.335
1.00
16.07
C


ATOM
861
CD
ARG
A
69
−69.378
26.578
−11.701
1.00
14.26
C


ATOM
864
NE
ARG
A
69
−68.915
25.205
−11.677
1.00
12.82
N


ATOM
866
CZ
ARG
A
69
−69.063
24.346
−12.676
1.00
12.42
C


ATOM
867
NH1
ARG
A
69
−69.676
24.694
−13.792
1.00
12.27
N


ATOM
870
NH2
ARG
A
69
−68.601
23.119
−12.553
1.00
13.23
N


ATOM
873
C
ARG
A
69
−66.010
29.409
−12.692
1.00
17.62
C


ATOM
874
O
ARG
A
69
−66.281
29.401
−13.892
1.00
17.10
O


ATOM
876
N
PHE
A
70
−65.811
30.529
−11.985
1.00
19.11
N


ATOM
877
CA
PHE
A
70
−65.936
31.891
−12.530
1.00
19.96
C


ATOM
879
CB
PHE
A
70
−67.024
32.666
−11.763
1.00
19.89
C


ATOM
882
CG
PHE
A
70
−68.365
32.010
−11.784
1.00
18.68
C


ATOM
883
CD1
PHE
A
70
−69.158
32.082
−12.903
1.00
18.05
C


ATOM
885
CE1
PHE
A
70
−70.376
31.452
−12.938
1.00
18.03
C


ATOM
887
CZ
PHE
A
70
−70.825
30.754
−11.851
1.00
17.26
C


ATOM
889
CE2
PHE
A
70
−70.051
30.672
−10.736
1.00
17.60
C


ATOM
891
CD2
PHE
A
70
−68.822
31.300
−10.700
1.00
17.86
C


ATOM
893
C
PHE
A
70
−64.627
32.661
−12.400
1.00
21.34
C


ATOM
894
O
PHE
A
70
−64.629
33.862
−12.171
1.00
21.25
O


ATOM
896
N
GLU
A
71
−63.501
31.977
−12.541
1.00
23.19
N


ATOM
897
CA
GLU
A
71
−62.211
32.591
−12.214
1.00
24.60
C


ATOM
899
CB
GLU
A
71
−61.127
31.524
−12.073
1.00
25.03
C


ATOM
902
CG
GLU
A
71
−59.717
32.059
−11.861
1.00
27.10
C


ATOM
905
CD
GLU
A
71
−58.712
30.949
−11.549
1.00
30.10
C


ATOM
906
OE1
GLU
A
71
−59.084
30.007
−10.796
1.00
32.05
O


ATOM
907
OE2
GLU
A
71
−57.558
31.027
−12.051
1.00
30.50
O


ATOM
908
C
GLU
A
71
−61.809
33.644
−13.241
1.00
25.23
C


ATOM
909
O
GLU
A
71
−61.362
34.717
−12.862
1.00
25.54
O


ATOM
911
N
SER
A
72
−61.979
33.345
−14.530
1.00
25.91
N


ATOM
912
CA
SER
A
72
−61.641
34.294
−15.588
1.00
26.37
C


ATOM
914
CB
SER
A
72
−61.656
33.606
−16.941
1.00
26.32
C


ATOM
917
OG
SER
A
72
−62.985
33.278
−17.291
1.00
27.28
O


ATOM
919
C
SER
A
72
−62.609
35.476
−15.608
1.00
26.84
C


ATOM
920
O
SER
A
72
−62.197
36.601
−15.852
1.00
27.02
O


ATOM
922
N
ASP
A
73
−63.893
35.214
−15.361
1.00
27.45
N


ATOM
923
CA
ASP
A
73
−64.895
36.277
−15.246
1.00
27.87
C


ATOM
925
CB
ASP
A
73
−66.289
35.705
−14.968
1.00
27.99
C


ATOM
928
CG
ASP
A
73
−66.842
34.913
−16.140
1.00
29.49
C


ATOM
929
OD1
ASP
A
73
−67.215
35.530
−17.155
1.00
30.90
O


ATOM
930
OD2
ASP
A
73
−66.916
33.667
−16.054
1.00
31.85
O


ATOM
931
C
ASP
A
73
−64.518
37.221
−14.121
1.00
27.86
C


ATOM
932
O
ASP
A
73
−64.598
38.435
−14.286
1.00
28.05
O


ATOM
934
N
ILE
A
74
−64.118
36.652
−12.985
1.00
27.88
N


ATOM
935
CA
ILE
A
74
−63.698
37.422
−11.821
1.00
27.98
C


ATOM
937
CB
ILE
A
74
−63.348
36.521
−10.637
1.00
27.71
C


ATOM
939
CG1
ILE
A
74
−64.607
36.007
−9.960
1.00
27.60
C


ATOM
942
CD1
ILE
A
74
−64.355
34.846
−9.023
1.00
27.67
C


ATOM
946
CG2
ILE
A
74
−62.551
37.272
−9.621
1.00
26.82
C


ATOM
950
C
ILE
A
74
−62.472
38.252
−12.133
1.00
28.80
C


ATOM
951
O
ILE
A
74
−62.475
39.453
−11.917
1.00
29.00
O


ATOM
953
N
ARG
A
75
−61.415
37.616
−12.628
1.00
29.80
N


ATOM
954
CA
ARG
A
75
−60.197
38.341
−12.968
1.00
30.79
C


ATOM
956
CB
ARG
A
75
−59.286
37.507
−13.841
1.00
31.33
C


ATOM
959
CG
ARG
A
75
−58.506
36.441
−13.115
1.00
34.25
C


ATOM
962
CD
ARG
A
75
−57.286
36.009
−13.929
1.00
37.53
C


ATOM
965
NE
ARG
A
75
−56.238
37.019
−13.799
1.00
40.89
N


ATOM
967
CZ
ARG
A
75
−54.934
36.807
−13.967
1.00
44.24
C


ATOM
968
NH1
ARG
A
75
−54.459
35.602
−14.294
1.00
45.02
N


ATOM
971
NH2
ARG
A
75
−54.089
37.821
−13.796
1.00
45.88
N


ATOM
974
C
ARG
A
75
−60.548
39.596
−13.727
1.00
30.95
C


ATOM
975
O
ARG
A
75
−60.163
40.685
−13.336
1.00
31.27
O


ATOM
977
N
ARG
A
76
−61.293
39.435
−14.815
1.00
31.31
N


ATOM
978
CA
ARG
A
76
−61.712
40.567
−15.638
1.00
31.62
C


ATOM
980
CB
ARG
A
76
−62.593
40.106
−16.794
1.00
31.97
C


ATOM
983
CG
ARG
A
76
−61.833
39.392
−17.895
1.00
32.91
C


ATOM
986
CD
ARG
A
76
−62.615
39.453
−19.205
1.00
34.43
C


ATOM
989
NE
ARG
A
76
−63.925
38.803
−19.128
1.00
35.66
N


ATOM
991
CZ
ARG
A
76
−64.115
37.480
−19.106
1.00
37.15
C


ATOM
992
NH1
ARG
A
76
−63.085
36.632
−19.129
1.00
37.93
N


ATOM
995
NH2
ARG
A
76
−65.347
36.992
−19.048
1.00
37.64
N


ATOM
998
C
ARG
A
76
−62.453
41.629
−14.849
1.00
31.41
C


ATOM
999
O
ARG
A
76
−62.144
42.798
−14.966
1.00
31.35
O


ATOM
1001
N
ALA
A
77
−63.437
41.230
−14.058
1.00
31.53
N


ATOM
1002
CA
ALA
A
77
−64.136
42.177
−13.211
1.00
31.73
C


ATOM
1004
CB
ALA
A
77
−65.074
41.468
−12.275
1.00
31.69
C


ATOM
1008
C
ALA
A
77
−63.107
42.950
−12.423
1.00
32.23
C


ATOM
1009
O
ALA
A
77
−63.116
44.176
−12.411
1.00
32.42
O


ATOM
1011
N
LEU
A
78
−62.198
42.223
−11.786
1.00
32.95
N


ATOM
1012
CA
LEU
A
78
−61.168
42.838
−10.963
1.00
33.51
C


ATOM
1014
CB
LEU
A
78
−60.319
41.775
−10.265
1.00
33.10
C


ATOM
1017
CG
LEU
A
78
−60.959
41.077
−9.081
1.00
32.30
C


ATOM
1019
CD1
LEU
A
78
−59.960
40.104
−8.472
1.00
30.80
C


ATOM
1023
CD2
LEU
A
78
−61.438
42.117
−8.056
1.00
31.89
C


ATOM
1027
C
LEU
A
78
−60.249
43.752
−11.753
1.00
34.70
C


ATOM
1028
O
LEU
A
78
−59.790
44.753
−11.214
1.00
35.18
O


ATOM
1030
N
ASP
A
79
−59.955
43.404
−13.007
1.00
35.91
N


ATOM
1031
CA
ASP
A
79
−59.054
44.205
−13.828
1.00
36.89
C


ATOM
1033
CB
ASP
A
79
−58.637
43.452
−15.083
1.00
37.06
C


ATOM
1036
CG
ASP
A
79
−57.518
44.150
−15.821
1.00
38.41
C


ATOM
1037
OD1
ASP
A
79
−57.800
45.080
−16.615
1.00
39.85
O


ATOM
1038
OD2
ASP
A
79
−56.347
43.775
−15.592
1.00
40.78
O


ATOM
1039
C
ASP
A
79
−59.721
45.505
−14.220
1.00
37.75
C


ATOM
1040
O
ASP
A
79
−59.105
46.568
−14.177
1.00
37.82
O


ATOM
1042
N
ARG
A
80
−60.984
45.401
−14.615
1.00
38.99
N


ATOM
1043
CA
ARG
A
80
−61.814
46.558
−14.923
1.00
39.88
C


ATOM
1045
CB
ARG
A
80
−63.191
46.106
−15.427
1.00
40.41
C


ATOM
1048
CG
ARG
A
80
−64.025
47.183
−16.122
1.00
42.77
C


ATOM
1051
CD
ARG
A
80
−65.347
46.624
−16.718
1.00
45.79
C


ATOM
1054
NE
ARG
A
80
−66.107
45.799
−15.765
1.00
48.55
N


ATOM
1056
CZ
ARG
A
80
−66.136
44.457
−15.742
1.00
50.37
C


ATOM
1057
NH1
ARG
A
80
−65.450
43.724
−16.636
1.00
49.78
N


ATOM
1060
NH2
ARG
A
80
−66.868
43.835
−14.805
1.00
50.77
N


ATOM
1063
C
ARG
A
80
−61.946
47.409
−13.668
1.00
39.80
C


ATOM
1064
O
ARG
A
80
−61.840
48.630
−13.742
1.00
40.37
O


ATOM
1066
N
PHE
A
81
−62.136
46.771
−12.516
1.00
39.62
N


ATOM
1067
CA
PHE
A
81
−62.216
47.505
−11.245
1.00
39.61
C


ATOM
1069
CB
PHE
A
81
−62.392
46.561
−10.053
1.00
39.74
C


ATOM
1072
CG
PHE
A
81
−62.282
47.247
−8.712
1.00
38.81
C


ATOM
1073
CD1
PHE
A
81
−63.238
48.153
−8.315
1.00
39.03
C


ATOM
1075
CE1
PHE
A
81
−63.153
48.787
−7.100
1.00
39.69
C


ATOM
1077
CZ
PHE
A
81
−62.095
48.515
−6.261
1.00
39.54
C


ATOM
1079
CE2
PHE
A
81
−61.134
47.608
−6.647
1.00
38.66
C


ATOM
1081
CD2
PHE
A
81
−61.228
46.986
−7.865
1.00
38.27
C


ATOM
1083
C
PHE
A
81
−61.006
48.378
−10.970
1.00
39.58
C


ATOM
1084
O
PHE
A
81
−61.165
49.535
−10.581
1.00
39.76
O


ATOM
1086
N
VAL
A
82
−59.810
47.817
−11.142
1.00
39.50
N


ATOM
1087
CA
VAL
A
82
−58.575
48.562
−10.892
1.00
39.47
C


ATOM
1089
CB
VAL
A
82
−57.314
47.659
−10.977
1.00
39.37
C


ATOM
1091
CG1
VAL
A
82
−56.108
48.426
−11.521
1.00
39.06
C


ATOM
1095
CG2
VAL
A
82
−57.009
47.079
−9.614
1.00
39.23
C


ATOM
1099
C
VAL
A
82
−58.457
49.755
−11.833
1.00
39.54
C


ATOM
1100
O
VAL
A
82
−58.222
50.884
−11.378
1.00
39.45
O


ATOM
1102
N
SER
A
83
−58.678
49.513
−13.126
1.00
39.65
N


ATOM
1103
CA
SER
A
83
−58.469
50.535
−14.157
1.00
39.82
C


ATOM
1105
CB
SER
A
83
−58.196
49.880
−15.526
1.00
39.84
C


ATOM
1108
OG
SER
A
83
−59.138
48.870
−15.829
1.00
40.00
O


ATOM
1110
C
SER
A
83
−59.617
51.561
−14.219
1.00
39.62
C


ATOM
1111
O
SER
A
83
−60.149
51.853
−15.286
1.00
39.59
O


ATOM
1113
N
SER
A
84
−59.957
52.108
−13.052
1.00
39.44
N


ATOM
1114
CA
SER
A
84
−60.926
53.197
−12.910
1.00
39.13
C


ATOM
1116
CB
SER
A
84
−62.238
52.834
−13.618
1.00
39.03
C


ATOM
1119
OG
SER
A
84
−62.620
51.505
−13.325
1.00
37.97
O


ATOM
1121
C
SER
A
84
−61.243
53.580
−11.446
1.00
39.17
C


ATOM
1122
O
SER
A
84
−62.189
54.341
−11.230
1.00
39.58
O


ATOM
1124
N
GLY
A
85
−60.482
53.077
−10.457
1.00
38.74
N


ATOM
1125
CA
GLY
A
85
−60.795
53.282
−9.024
1.00
38.26
C


ATOM
1128
C
GLY
A
85
−61.724
52.238
−8.413
1.00
37.79
C


ATOM
1129
O
GLY
A
85
−61.529
51.799
−7.273
1.00
36.84
O


ATOM
1131
N
SER
A
94
−63.557
54.187
1.994
1.00
28.27
N


ATOM
1132
CA
SER
A
94
−64.069
52.892
2.476
1.00
28.03
C


ATOM
1134
CB
SER
A
94
−64.856
52.225
1.359
1.00
27.84
C


ATOM
1137
OG
SER
A
94
−65.234
50.921
1.743
1.00
28.20
O


ATOM
1139
C
SER
A
94
−62.992
51.910
2.934
1.00
27.83
C


ATOM
1140
O
SER
A
94
−62.339
51.325
2.090
1.00
28.15
O


ATOM
1142
N
LEU
A
95
−62.815
51.705
4.245
1.00
27.70
N


ATOM
1143
CA
LEU
A
95
−61.813
50.728
4.751
1.00
27.59
C


ATOM
1145
CB
LEU
A
95
−61.692
50.747
6.282
1.00
27.49
C


ATOM
1148
CG
LEU
A
95
−60.871
49.596
6.909
1.00
27.47
C


ATOM
1150
CD1
LEU
A
95
−59.400
49.900
6.902
1.00
27.70
C


ATOM
1154
CD2
LEU
A
95
−61.292
49.274
8.329
1.00
27.50
C


ATOM
1158
C
LEU
A
95
−62.173
49.314
4.318
1.00
27.71
C


ATOM
1159
O
LEU
A
95
−61.303
48.537
3.914
1.00
27.73
O


ATOM
1161
N
HIS
A
96
−63.461
48.989
4.437
1.00
27.74
N


ATOM
1162
CA
HIS
A
96
−64.003
47.715
3.977
1.00
27.54
C


ATOM
1164
CB
HIS
A
96
−65.497
47.680
4.230
1.00
27.57
C


ATOM
1167
CG
HIS
A
96
−66.161
46.482
3.654
1.00
28.65
C


ATOM
1168
ND1
HIS
A
96
−65.790
45.201
3.988
1.00
30.59
N


ATOM
1170
CE1
HIS
A
96
−66.540
44.341
3.325
1.00
31.88
C


ATOM
1172
NE2
HIS
A
96
−67.384
45.021
2.570
1.00
32.06
N


ATOM
1174
CD2
HIS
A
96
−67.166
46.364
2.758
1.00
30.47
C


ATOM
1176
C
HIS
A
96
−63.705
47.450
2.492
1.00
27.09
C


ATOM
1177
O
HIS
A
96
−63.128
46.425
2.150
1.00
27.18
O


ATOM
1179
N
GLY
A
97
−64.079
48.386
1.625
1.00
26.55
N


ATOM
1180
CA
GLY
A
97
−63.760
48.300
.205
1.00
26.26
C


ATOM
1183
C
GLY
A
97
−62.277
48.112
−.088
1.00
26.23
C


ATOM
1184
O
GLY
A
97
−61.918
47.334
−.974
1.00
26.39
O


ATOM
1186
N
THR
A
98
−61.415
48.815
.653
1.00
25.83
N


ATOM
1187
CA
THR
A
98
−59.967
48.755
.435
1.00
25.36
C


ATOM
1189
CB
THR
A
98
−59.204
49.847
1.215
1.00
25.07
C


ATOM
1191
OG1
THR
A
98
−59.796
51.123
.980
1.00
23.62
O


ATOM
1193
CG2
THR
A
98
−57.755
49.905
.780
1.00
24.88
C


ATOM
1197
C
THR
A
98
−59.416
47.389
.845
1.00
25.69
C


ATOM
1198
O
THR
A
98
−58.622
46.802
.111
1.00
25.66
O


ATOM
1200
N
ALA
A
99
−59.847
46.890
2.008
1.00
25.95
N


ATOM
1201
CA
ALA
A
99
−59.468
45.540
2.495
1.00
25.93
C


ATOM
1203
CB
ALA
A
99
−59.945
45.330
3.929
1.00
25.82
C


ATOM
1207
C
ALA
A
99
−59.984
44.404
1.595
1.00
25.64
C


ATOM
1208
O
ALA
A
99
−59.204
43.599
1.102
1.00
25.38
O


ATOM
1210
N
LEU
A
100
−61.290
44.348
1.371
1.00
25.46
N


ATOM
1211
CA
LEU
A
100
−61.856
43.335
.474
1.00
25.55
C


ATOM
1213
CB
LEU
A
100
−63.361
43.540
.326
1.00
25.37
C


ATOM
1216
CG
LEU
A
100
−64.118
42.464
−.443
1.00
25.41
C


ATOM
1218
CD1
LEU
A
100
−64.038
41.115
.281
1.00
26.01
C


ATOM
1222
CD2
LEU
A
100
−65.555
42.893
−.639
1.00
25.19
C


ATOM
1226
C
LEU
A
100
−61.196
43.353
−.919
1.00
25.70
C


ATOM
1227
O
LEU
A
100
−60.819
42.301
−1.469
1.00
25.36
O


ATOM
1229
N
SER
A
101
−61.064
44.550
−1.489
1.00
25.77
N


ATOM
1230
CA
SER
A
101
−60.434
44.688
−2.805
1.00
25.67
C


ATOM
1232
CB
SER
A
101
−60.629
46.098
−3.375
1.00
25.83
C


ATOM
1235
OG
SER
A
101
−59.840
47.064
−2.688
1.00
26.98
O


ATOM
1237
C
SER
A
101
−58.947
44.336
−2.756
1.00
25.01
C


ATOM
1238
O
SER
A
101
−58.427
43.705
−3.672
1.00
24.94
O


ATOM
1240
N
PHE
A
102
−58.270
44.737
−1.687
1.00
24.30
N


ATOM
1241
CA
PHE
A
102
−56.852
44.436
−1.560
1.00
23.99
C


ATOM
1243
CB
PHE
A
102
−56.295
44.963
−.229
1.00
23.84
C


ATOM
1246
CG
PHE
A
102
−54.860
44.606
.027
1.00
23.24
C


ATOM
1247
CD1
PHE
A
102
−53.849
45.504
−.259
1.00
23.56
C


ATOM
1249
CE1
PHE
A
102
−52.507
45.182
−.007
1.00
24.50
C


ATOM
1251
CZ
PHE
A
102
−52.175
43.941
.540
1.00
24.27
C


ATOM
1253
CE2
PHE
A
102
−53.180
43.042
.834
1.00
23.95
C


ATOM
1255
CD2
PHE
A
102
−54.520
43.376
.576
1.00
23.45
C


ATOM
1257
C
PHE
A
102
−56.673
42.928
−1.679
1.00
23.65
C


ATOM
1258
O
PHE
A
102
−55.890
42.442
−2.501
1.00
23.55
O


ATOM
1260
N
ARG
A
103
−57.443
42.202
−.880
1.00
23.20
N


ATOM
1261
CA
ARG
A
103
−57.298
40.765
−.764
1.00
22.84
C


ATOM
1263
CB
ARG
A
103
−58.250
40.231
.291
1.00
23.01
C


ATOM
1266
CG
ARG
A
103
−58.054
38.774
.597
1.00
23.93
C


ATOM
1269
CD
ARG
A
103
−58.703
38.376
1.937
1.00
23.84
C


ATOM
1272
NE
ARG
A
103
−60.150
38.538
1.919
1.00
22.36
N


ATOM
1274
CZ
ARG
A
103
−60.982
37.763
1.240
1.00
20.62
C


ATOM
1275
NH1
ARG
A
103
−60.512
36.773
.490
1.00
20.19
N


ATOM
1278
NH2
ARG
A
103
−62.288
38.003
1.293
1.00
20.32
N


ATOM
1281
C
ARG
A
103
−57.602
40.117
−2.082
1.00
22.34
C


ATOM
1282
O
ARG
A
103
−56.826
39.303
−2.576
1.00
22.37
O


ATOM
1284
N
LEU
A
104
−58.729
40.496
−2.667
1.00
21.98
N


ATOM
1285
CA
LEU
A
104
−59.129
39.926
−3.951
1.00
21.60
C


ATOM
1287
CB
LEU
A
104
−60.503
40.442
−4.378
1.00
21.23
C


ATOM
1290
CG
LEU
A
104
−61.590
39.832
−3.522
1.00
20.30
C


ATOM
1292
CD1
LEU
A
104
−62.949
40.401
−3.856
1.00
19.53
C


ATOM
1296
CD2
LEU
A
104
−61.554
38.344
−3.725
1.00
19.71
C


ATOM
1300
C
LEU
A
104
−58.101
40.198
−5.040
1.00
21.48
C


ATOM
1301
O
LEU
A
104
−57.743
39.294
−5.780
1.00
21.62
O


ATOM
1303
N
LEU
A
105
−57.623
41.432
−5.139
1.00
21.14
N


ATOM
1304
CA
LEU
A
105
−56.655
41.747
−6.173
1.00
21.05
C


ATOM
1306
CB
LEU
A
105
−56.352
43.248
−6.205
1.00
20.94
C


ATOM
1309
CG
LEU
A
105
−57.465
44.165
−6.732
1.00
20.06
C


ATOM
1311
CD1
LEU
A
105
−57.060
45.617
−6.580
1.00
17.92
C


ATOM
1315
CD2
LEU
A
105
−57.804
43.857
−8.174
1.00
19.18
C


ATOM
1319
C
LEU
A
105
−55.372
40.908
−5.997
1.00
21.32
C


ATOM
1320
O
LEU
A
105
−54.840
40.357
−6.976
1.00
21.37
O


ATOM
1322
N
ARG
A
106
−54.893
40.777
−4.763
1.00
21.25
N


ATOM
1323
CA
ARG
A
106
−53.678
40.003
−4.528
1.00
21.33
C


ATOM
1325
CB
ARG
A
106
−53.151
40.211
−3.117
1.00
21.53
C


ATOM
1328
CG
ARG
A
106
−51.772
39.588
−2.917
1.00
22.60
C


ATOM
1331
CD
ARG
A
106
−51.098
40.091
−1.666
1.00
23.60
C


ATOM
1334
NE
ARG
A
106
−50.374
41.330
−1.897
1.00
24.60
N


ATOM
1336
CZ
ARG
A
106
−49.606
41.916
−.989
1.00
26.54
C


ATOM
1337
NH1
ARG
A
106
−49.469
41.380
.225
1.00
26.98
N


ATOM
1340
NH2
ARG
A
106
−48.967
43.041
−1.295
1.00
27.19
N


ATOM
1343
C
ARG
A
106
−53.888
38.517
−4.767
1.00
21.19
C


ATOM
1344
O
ARG
A
106
−52.998
37.821
−5.278
1.00
21.18
O


ATOM
1346
N
GLN
A
107
−55.061
38.028
−4.378
1.00
21.09
N


ATOM
1347
CA
GLN
A
107
−55.425
36.635
−4.618
1.00
20.76
C


ATOM
1349
CB
GLN
A
107
−56.861
36.378
−4.168
1.00
20.51
C


ATOM
1352
CG
GLN
A
107
−57.329
34.941
−4.327
1.00
19.68
C


ATOM
1355
CD
GLN
A
107
−58.790
34.772
−4.010
1.00
18.26
C


ATOM
1356
OE1
GLN
A
107
−59.395
35.591
−3.334
1.00
17.44
O


ATOM
1357
NE2
GLN
A
107
−59.366
33.698
−4.497
1.00
18.48
N


ATOM
1360
C
GLN
A
107
−55.304
36.306
−6.094
1.00
20.93
C


ATOM
1361
O
GLN
A
107
−54.917
35.212
−6.447
1.00
21.45
O


ATOM
1363
N
HIS
A
108
−55.642
37.260
−6.951
1.00
21.09
N


ATOM
1364
CA
HIS
A
108
−55.686
37.030
−8.379
1.00
21.28
C


ATOM
1366
CB
HIS
A
108
−57.024
37.541
−8.913
1.00
21.02
C


ATOM
1369
CG
HIS
A
108
−58.182
36.706
−8.478
1.00
19.70
C


ATOM
1370
ND1
HIS
A
108
−58.637
35.632
−9.210
1.00
19.91
N


ATOM
1372
CE1
HIS
A
108
−59.641
35.060
−8.571
1.00
19.27
C


ATOM
1374
NE2
HIS
A
108
−59.851
35.720
−7.449
1.00
18.10
N


ATOM
1376
CD2
HIS
A
108
−58.950
36.753
−7.367
1.00
18.98
C


ATOM
1378
C
HIS
A
108
−54.456
37.621
−9.108
1.00
22.21
C


ATOM
1379
O
HIS
A
108
−54.505
37.980
−10.304
1.00
21.80
O


ATOM
1381
N
GLY
A
109
−53.345
37.686
−8.375
1.00
23.19
N


ATOM
1382
CA
GLY
A
109
−52.055
38.007
−8.956
1.00
24.17
C


ATOM
1385
C
GLY
A
109
−51.813
39.470
−9.275
1.00
25.25
C


ATOM
1386
O
GLY
A
109
−50.767
39.802
−9.845
1.00
25.26
O


ATOM
1388
N
PHE
A
110
−52.755
40.349
−8.917
1.00
26.42
N


ATOM
1389
CA
PHE
A
110
−52.575
41.783
−9.154
1.00
27.33
C


ATOM
1391
CB
PHE
A
110
−53.895
42.546
−9.067
1.00
27.36
C


ATOM
1394
CG
PHE
A
110
−54.838
42.250
−10.193
1.00
27.83
C


ATOM
1395
CD1
PHE
A
110
−54.633
42.801
−11.439
1.00
28.69
C


ATOM
1397
CE1
PHE
A
110
−55.490
42.529
−12.488
1.00
28.91
C


ATOM
1399
CZ
PHE
A
110
−56.568
41.702
−12.295
1.00
28.85
C


ATOM
1401
CE2
PHE
A
110
−56.787
41.146
−11.060
1.00
28.67
C


ATOM
1403
CD2
PHE
A
110
−55.923
41.418
−10.013
1.00
28.39
C


ATOM
1405
C
PHE
A
110
−51.582
42.369
−8.163
1.00
28.25
C


ATOM
1406
O
PHE
A
110
−51.449
41.890
−7.024
1.00
28.71
O


ATOM
1408
N
GLU
A
111
−50.886
43.411
−8.607
1.00
29.05
N


ATOM
1409
CA
GLU
A
111
−49.951
44.142
−7.760
1.00
29.66
C


ATOM
1411
CB
GLU
A
111
−48.902
44.803
−8.648
1.00
30.37
C


ATOM
1414
CG
GLU
A
111
−47.750
45.509
−7.947
1.00
33.30
C


ATOM
1417
CD
GLU
A
111
−46.916
46.342
−8.946
1.00
38.00
C


ATOM
1418
OE1
GLU
A
111
−46.312
45.750
−9.885
1.00
39.60
O


ATOM
1419
OE2
GLU
A
111
−46.886
47.594
−8.802
1.00
40.87
O


ATOM
1420
C
GLU
A
111
−50.724
45.184
−6.935
1.00
28.98
C


ATOM
1421
O
GLU
A
111
−51.293
46.123
−7.495
1.00
28.45
O


ATOM
1423
N
VAL
A
112
−50.787
44.961
−5.618
1.00
28.47
N


ATOM
1424
CA
VAL
A
112
−51.284
45.950
−4.655
1.00
28.09
C


ATOM
1426
CB
VAL
A
112
−52.564
45.501
−3.908
1.00
27.76
C


ATOM
1428
CG1
VAL
A
112
−53.745
45.466
−4.836
1.00
27.38
C


ATOM
1432
CG2
VAL
A
112
−52.360
44.167
−3.252
1.00
27.80
C


ATOM
1436
C
VAL
A
112
−50.193
46.199
−3.631
1.00
28.15
C


ATOM
1437
O
VAL
A
112
−49.339
45.354
−3.436
1.00
27.90
O


ATOM
1439
N
SER
A
113
−50.241
47.354
−2.974
1.00
28.54
N


ATOM
1440
CA
SER
A
113
−49.194
47.782
−2.047
1.00
28.93
C


ATOM
1442
CB
SER
A
113
−48.563
49.064
−2.564
1.00
29.12
C


ATOM
1445
OG
SER
A
113
−47.692
49.644
−1.619
1.00
29.57
O


ATOM
1447
C
SER
A
113
−49.780
48.041
−.685
1.00
29.13
C


ATOM
1448
O
SER
A
113
−50.937
48.426
−.581
1.00
28.90
O


ATOM
1450
N
GLN
A
114
−48.985
47.857
.367
1.00
29.84
N


ATOM
1451
CA
GLN
A
114
−49.524
48.019
1.736
1.00
30.40
C


ATOM
1453
CB
GLN
A
114
−48.568
47.507
2.830
1.00
30.34
C


ATOM
1456
CG
GLN
A
114
−47.199
48.159
2.844
1.00
30.61
C


ATOM
1459
CD
GLN
A
114
−46.456
47.928
4.147
1.00
30.77
C


ATOM
1460
OE1
GLN
A
114
−46.876
47.129
4.982
1.00
31.92
O


ATOM
1461
NE2
GLN
A
114
−45.341
48.624
4.324
1.00
30.02
N


ATOM
1464
C
GLN
A
114
−49.939
49.440
2.068
1.00
30.62
C


ATOM
1465
O
GLN
A
114
−50.573
49.644
3.094
1.00
30.73
O


ATOM
1467
N
GLU
A
115
−49.592
50.407
1.216
1.00
30.94
N


ATOM
1468
CA
GLU
A
115
−49.963
51.794
1.458
1.00
31.45
C


ATOM
1470
CB
GLU
A
115
−48.881
52.777
.975
1.00
31.86
C


ATOM
1473
CG
GLU
A
115
−48.489
52.723
−.509
1.00
33.48
C


ATOM
1476
CD
GLU
A
115
−47.006
53.100
−.759
1.00
36.13
C


ATOM
1477
OE1
GLU
A
115
−46.106
52.668
.022
1.00
38.72
O


ATOM
1478
OE2
GLU
A
115
−46.741
53.811
−1.754
1.00
35.70
O


ATOM
1479
C
GLU
A
115
−51.338
52.125
.895
1.00
31.45
C


ATOM
1480
O
GLU
A
115
−51.716
53.283
.808
1.00
31.80
O


ATOM
1482
N
ALA
A
116
−52.108
51.105
.549
1.00
31.58
N


ATOM
1483
CA
ALA
A
116
−53.524
51.286
.284
1.00
31.67
C


ATOM
1485
CB
ALA
A
116
−54.071
50.092
−.461
1.00
31.62
C


ATOM
1489
C
ALA
A
116
−54.273
51.473
1.596
1.00
31.88
C


ATOM
1490
O
ALA
A
116
−55.428
51.874
1.604
1.00
31.82
O


ATOM
1492
N
PHE
A
117
−53.608
51.161
2.702
1.00
32.39
N


ATOM
1493
CA
PHE
A
117
−54.193
51.271
4.030
1.00
32.87
C


ATOM
1495
CB
PHE
A
117
−53.884
50.003
4.856
1.00
32.79
C


ATOM
1498
CG
PHE
A
117
−54.539
48.758
4.323
1.00
31.03
C


ATOM
1499
CD1
PHE
A
117
−53.782
47.749
3.760
1.00
28.79
C


ATOM
1501
CE1
PHE
A
117
−54.379
46.620
3.265
1.00
28.36
C


ATOM
1503
CZ
PHE
A
117
−55.754
46.489
3.316
1.00
29.50
C


ATOM
1505
CE2
PHE
A
117
−56.527
47.493
3.872
1.00
29.91
C


ATOM
1507
CD2
PHE
A
117
−55.917
48.613
4.375
1.00
30.09
C


ATOM
1509
C
PHE
A
117
−53.683
52.493
4.773
1.00
33.71
C


ATOM
1510
O
PHE
A
117
−54.012
52.678
5.937
1.00
33.61
O


ATOM
1512
N
SER
A
118
−52.886
53.325
4.107
1.00
35.09
N


ATOM
1513
CA
SER
A
118
−52.186
54.429
4.779
1.00
36.12
C


ATOM
1515
CB
SER
A
118
−51.073
54.990
3.895
1.00
36.09
C


ATOM
1518
OG
SER
A
118
−51.607
55.747
2.820
1.00
35.96
O


ATOM
1520
C
SER
A
118
−53.134
55.552
5.184
1.00
37.17
C


ATOM
1521
O
SER
A
118
−52.873
56.255
6.166
1.00
37.22
O


ATOM
1523
N
GLY
A
119
−54.231
55.705
4.436
1.00
38.40
N


ATOM
1524
CA
GLY
A
119
−55.227
56.735
4.708
1.00
39.47
C


ATOM
1527
C
GLY
A
119
−56.255
56.398
5.778
1.00
40.62
C


ATOM
1528
O
GLY
A
119
−57.259
57.086
5.877
1.00
40.74
O


ATOM
1530
N
PHE
A
120
−56.024
55.344
6.563
1.00
42.13
N


ATOM
1531
CA
PHE
A
120
−56.906
54.971
7.684
1.00
43.27
C


ATOM
1533
CB
PHE
A
120
−57.510
53.586
7.455
1.00
43.21
C


ATOM
1536
CG
PHE
A
120
−58.176
53.439
6.134
1.00
42.36
C


ATOM
1537
CD1
PHE
A
120
−59.418
54.006
5.911
1.00
41.23
C


ATOM
1539
CE1
PHE
A
120
−60.035
53.880
4.691
1.00
40.87
C


ATOM
1541
CZ
PHE
A
120
−59.414
53.182
3.675
1.00
41.03
C


ATOM
1543
CE2
PHE
A
120
−58.167
52.618
3.883
1.00
40.92
C


ATOM
1545
CD2
PHE
A
120
−57.556
52.746
5.106
1.00
41.30
C


ATOM
1547
C
PHE
A
120
−56.179
54.952
9.019
1.00
44.71
C


ATOM
1548
O
PHE
A
120
−56.732
54.489
10.025
1.00
44.80
O


ATOM
1550
N
LYS
A
121
−54.939
55.430
9.021
1.00
46.38
N


ATOM
1551
CA
LYS
A
121
−54.153
55.519
10.235
1.00
47.81
C


ATOM
1553
CB
LYS
A
121
−52.722
55.050
9.957
1.00
47.86
C


ATOM
1556
CG
LYS
A
121
−52.648
53.537
9.648
1.00
48.24
C


ATOM
1559
CD
LYS
A
121
−51.417
53.119
8.823
1.00
49.41
C


ATOM
1562
CE
LYS
A
121
−50.095
53.142
9.621
1.00
49.88
C


ATOM
1565
NZ
LYS
A
121
−49.944
52.005
10.584
1.00
49.89
N


ATOM
1569
C
LYS
A
121
−54.257
56.964
10.735
1.00
49.00
C


ATOM
1570
O
LYS
A
121
−54.712
57.839
10.000
1.00
49.11
O


ATOM
1572
N
ASP
A
122
−53.899
57.206
11.995
1.00
50.51
N


ATOM
1573
CA
ASP
A
122
−54.078
58.537
12.596
1.00
51.46
C


ATOM
1575
CB
ASP
A
122
−54.604
58.443
14.050
1.00
51.29
C


ATOM
1578
CG
ASP
A
122
−53.570
57.919
15.036
1.00
50.73
C


ATOM
1579
OD1
ASP
A
122
−52.353
58.025
14.777
1.00
49.90
O


ATOM
1580
OD2
ASP
A
122
−53.988
57.398
16.088
1.00
49.66
O


ATOM
1581
C
ASP
A
122
−52.792
59.358
12.502
1.00
52.60
C


ATOM
1582
O
ASP
A
122
−51.830
58.953
11.835
1.00
52.65
O


ATOM
1584
N
GLN
A
123
−52.802
60.512
13.172
1.00
53.88
N


ATOM
1585
CA
GLN
A
123
−51.680
61.456
13.195
1.00
54.66
C


ATOM
1587
CB
GLN
A
123
−51.993
62.594
14.178
1.00
55.01
C


ATOM
1590
CG
GLN
A
123
−53.249
63.442
13.831
1.00
56.15
C


ATOM
1593
CD
GLN
A
123
−52.943
64.736
13.064
1.00
57.64
C


ATOM
1594
OE1
GLN
A
123
−51.779
65.102
12.859
1.00
59.10
O


ATOM
1595
NE2
GLN
A
123
−54.001
65.438
12.652
1.00
57.63
N


ATOM
1598
C
GLN
A
123
−50.365
60.772
13.591
1.00
54.87
C


ATOM
1599
O
GLN
A
123
−49.309
61.078
13.034
1.00
54.67
O


ATOM
1601
N
ASN
A
124
−50.458
59.834
14.538
1.00
55.21
N


ATOM
1602
CA
ASN
A
124
−49.300
59.137
15.109
1.00
55.39
C


ATOM
1604
CB
ASN
A
124
−49.421
59.122
16.637
1.00
55.52
C


ATOM
1607
CG
ASN
A
124
−49.833
60.478
17.202
1.00
55.98
C


ATOM
1608
OD1
ASN
A
124
−49.093
61.460
17.091
1.00
56.93
O


ATOM
1609
ND2
ASN
A
124
−51.026
60.540
17.797
1.00
55.99
N


ATOM
1612
C
ASN
A
124
−49.116
57.705
14.589
1.00
55.27
C


ATOM
1613
O
ASN
A
124
−48.530
56.864
15.271
1.00
55.18
O


ATOM
1615
N
GLY
A
125
−49.626
57.430
13.391
1.00
55.18
N


ATOM
1616
CA
GLY
A
125
−49.311
56.190
12.672
1.00
55.08
C


ATOM
1619
C
GLY
A
125
−49.994
54.897
13.119
1.00
54.90
C


ATOM
1620
O
GLY
A
125
−49.543
53.806
12.750
1.00
55.24
O


ATOM
1622
N
ASN
A
126
−51.072
55.008
13.900
1.00
54.23
N


ATOM
1623
CA
ASN
A
126
−51.871
53.853
14.329
1.00
53.39
C


ATOM
1625
CB
ASN
A
126
−51.927
53.782
15.852
1.00
53.27
C


ATOM
1628
CG
ASN
A
126
−50.596
53.425
16.466
1.00
52.81
C


ATOM
1629
OD1
ASN
A
126
−49.636
53.113
15.764
1.00
52.29
O


ATOM
1630
ND2
ASN
A
126
−50.532
53.460
17.789
1.00
52.53
N


ATOM
1633
C
ASN
A
126
−53.274
53.978
13.771
1.00
52.82
C


ATOM
1634
O
ASN
A
126
−53.724
55.081
13.494
1.00
52.69
O


ATOM
1636
N
PHE
A
127
−53.968
52.857
13.607
1.00
52.17
N


ATOM
1637
CA
PHE
A
127
−55.287
52.875
12.962
1.00
51.56
C


ATOM
1639
CB
PHE
A
127
−55.805
51.451
12.721
1.00
51.35
C


ATOM
1642
CG
PHE
A
127
−55.119
50.730
11.587
1.00
50.06
C


ATOM
1643
CD1
PHE
A
127
−54.068
49.856
11.831
1.00
48.90
C


ATOM
1645
CE1
PHE
A
127
−53.437
49.187
10.789
1.00
47.59
C


ATOM
1647
CZ
PHE
A
127
−53.857
49.383
9.495
1.00
47.40
C


ATOM
1649
CE2
PHE
A
127
−54.906
50.244
9.234
1.00
47.96
C


ATOM
1651
CD2
PHE
A
127
−55.533
50.915
10.276
1.00
48.85
C


ATOM
1653
C
PHE
A
127
−56.303
53.690
13.776
1.00
51.35
C


ATOM
1654
O
PHE
A
127
−56.347
53.594
15.002
1.00
51.09
O


ATOM
1656
N
LEU
A
128
−57.100
54.500
13.083
1.00
51.15
N


ATOM
1657
CA
LEU
A
128
−58.143
55.280
13.727
1.00
51.09
C


ATOM
1659
CB
LEU
A
128
−59.039
55.979
12.694
1.00
51.17
C


ATOM
1662
CG
LEU
A
128
−58.463
57.046
11.746
1.00
51.31
C


ATOM
1664
CD1
LEU
A
128
−59.571
57.628
10.855
1.00
50.76
C


ATOM
1668
CD2
LEU
A
128
−57.762
58.157
12.508
1.00
50.98
C


ATOM
1672
C
LEU
A
128
−58.996
54.353
14.581
1.00
51.06
C


ATOM
1673
O
LEU
A
128
−59.573
53.393
14.078
1.00
50.94
O


ATOM
1675
N
GLU
A
129
−59.053
54.637
15.879
1.00
51.12
N


ATOM
1676
CA
GLU
A
129
−59.916
53.913
16.808
1.00
50.99
C


ATOM
1678
CB
GLU
A
129
−60.019
54.665
18.132
1.00
51.14
C


ATOM
1681
CG
GLU
A
129
−58.850
54.431
19.049
1.00
52.04
C


ATOM
1684
CD
GLU
A
129
−58.903
53.069
19.697
1.00
53.22
C


ATOM
1685
OE1
GLU
A
129
−58.040
52.219
19.376
1.00
54.19
O


ATOM
1686
OE2
GLU
A
129
−59.823
52.848
20.516
1.00
53.51
O


ATOM
1687
C
GLU
A
129
−61.314
53.712
16.260
1.00
50.66
C


ATOM
1688
O
GLU
A
129
−61.791
52.587
16.200
1.00
50.83
O


ATOM
1690
N
ASN
A
130
−61.960
54.799
15.839
1.00
50.28
N


ATOM
1691
CA
ASN
A
130
−63.389
54.765
15.486
1.00
50.03
C


ATOM
1693
CB
ASN
A
130
−63.910
56.182
15.193
1.00
50.09
C


ATOM
1696
CG
ASN
A
130
−63.286
56.805
13.960
1.00
50.37
C


ATOM
1697
OD1
ASN
A
130
−62.504
56.177
13.249
1.00
51.44
O


ATOM
1698
ND2
ASN
A
130
−63.634
58.056
13.701
1.00
50.27
N


ATOM
1701
C
ASN
A
130
−63.804
53.782
14.369
1.00
49.56
C


ATOM
1702
O
ASN
A
130
−64.993
53.581
14.131
1.00
49.44
O


ATOM
1704
N
LEU
A
131
−62.828
53.175
13.699
1.00
49.21
N


ATOM
1705
CA
LEU
A
131
−63.087
52.103
12.729
1.00
48.78
C


ATOM
1707
CB
LEU
A
131
−61.846
51.832
11.875
1.00
48.64
C


ATOM
1710
CG
LEU
A
131
−61.445
52.939
10.898
1.00
48.26
C


ATOM
1712
CD1
LEU
A
131
−60.029
52.709
10.375
1.00
47.76
C


ATOM
1716
CD2
LEU
A
131
−62.441
53.039
9.754
1.00
47.32
C


ATOM
1720
C
LEU
A
131
−63.516
50.798
13.394
1.00
48.58
C


ATOM
1721
O
LEU
A
131
−64.093
49.939
12.731
1.00
48.64
O


ATOM
1723
N
LYS
A
132
−63.225
50.645
14.689
1.00
48.23
N


ATOM
1724
CA
LYS
A
132
−63.650
49.474
15.465
1.00
47.80
C


ATOM
1726
CB
LYS
A
132
−63.156
49.587
16.913
1.00
47.71
C


ATOM
1729
CG
LYS
A
132
−63.930
50.615
17.728
1.00
47.80
C


ATOM
1732
CD
LYS
A
132
−63.430
50.779
19.153
1.00
47.75
C


ATOM
1735
CE
LYS
A
132
−64.438
51.577
19.982
1.00
47.22
C


ATOM
1738
NZ
LYS
A
132
−63.769
52.364
21.033
1.00
46.65
N


ATOM
1742
C
LYS
A
132
−65.182
49.308
15.446
1.00
47.56
C


ATOM
1743
O
LYS
A
132
−65.696
48.204
15.616
1.00
47.76
O


ATOM
1745
N
GLU
A
133
−65.901
50.413
15.249
1.00
47.18
N


ATOM
1746
CA
GLU
A
133
−67.371
50.412
15.191
1.00
46.79
C


ATOM
1748
CB
GLU
A
133
−67.898
51.850
15.337
1.00
46.95
C


ATOM
1751
CG
GLU
A
133
−67.840
52.376
16.776
1.00
47.56
C


ATOM
1754
CD
GLU
A
133
−67.613
53.886
16.863
1.00
48.41
C


ATOM
1755
OE1
GLU
A
133
−68.247
54.648
16.093
1.00
47.67
O


ATOM
1756
OE2
GLU
A
133
−66.798
54.304
17.721
1.00
48.92
O


ATOM
1757
C
GLU
A
133
−67.949
49.770
13.918
1.00
46.10
C


ATOM
1758
O
GLU
A
133
−69.140
49.446
13.874
1.00
46.11
O


ATOM
1760
N
ASP
A
134
−67.103
49.601
12.896
1.00
45.11
N


ATOM
1761
CA
ASP
A
134
−67.477
48.995
11.608
1.00
44.06
C


ATOM
1763
CB
ASP
A
134
−66.934
49.868
10.474
1.00
43.97
C


ATOM
1766
CG
ASP
A
134
−67.380
49.411
9.104
1.00
44.08
C


ATOM
1767
OD1
ASP
A
134
−68.072
48.373
8.972
1.00
43.42
O


ATOM
1768
OD2
ASP
A
134
−67.020
50.116
8.141
1.00
44.84
O


ATOM
1769
C
ASP
A
134
−66.910
47.570
11.526
1.00
43.12
C


ATOM
1770
O
ASP
A
134
−65.788
47.357
11.080
1.00
43.19
O


ATOM
1772
N
ILE
A
135
−67.708
46.594
11.944
1.00
41.85
N


ATOM
1773
CA
ILE
A
135
−67.216
45.247
12.216
1.00
40.75
C


ATOM
1775
CB
ILE
A
135
−68.183
44.507
13.170
1.00
40.80
C


ATOM
1777
CG1
ILE
A
135
−68.227
45.226
14.524
1.00
41.21
C


ATOM
1780
CD1
ILE
A
135
−69.353
46.283
14.633
1.00
42.86
C


ATOM
1784
CG2
ILE
A
135
−67.776
43.063
13.373
1.00
40.88
C


ATOM
1788
C
ILE
A
135
−66.980
44.457
10.933
1.00
39.67
C


ATOM
1789
O
ILE
A
135
−66.166
43.537
10.901
1.00
39.37
O


ATOM
1791
N
LYS
A
136
−67.690
44.828
9.877
1.00
38.62
N


ATOM
1792
CA
LYS
A
136
−67.465
44.256
8.554
1.00
37.88
C


ATOM
1794
CB
LYS
A
136
−68.525
44.749
7.561
1.00
38.33
C


ATOM
1797
CG
LYS
A
136
−69.955
44.342
7.879
1.00
40.17
C


ATOM
1800
CD
LYS
A
136
−70.486
43.262
6.911
1.00
42.97
C


ATOM
1803
CE
LYS
A
136
−71.624
42.389
7.536
1.00
44.00
C


ATOM
1806
NZ
LYS
A
136
−72.950
43.086
7.726
1.00
43.82
N


ATOM
1810
C
LYS
A
136
−66.097
44.687
8.047
1.00
36.38
C


ATOM
1811
O
LYS
A
136
−65.399
43.914
7.388
1.00
36.24
O


ATOM
1813
N
ALA
A
137
−65.728
45.931
8.341
1.00
34.51
N


ATOM
1814
CA
ALA
A
137
−64.463
46.474
7.876
1.00
33.41
C


ATOM
1816
CB
ALA
A
137
−64.432
47.981
8.058
1.00
33.43
C


ATOM
1820
C
ALA
A
137
−63.291
45.827
8.600
1.00
32.30
C


ATOM
1821
O
ALA
A
137
−62.288
45.481
7.995
1.00
31.89
O


ATOM
1823
N
ILE
A
138
−63.437
45.662
9.905
1.00
31.41
N


ATOM
1824
CA
ILE
A
138
−62.395
45.074
10.737
1.00
30.60
C


ATOM
1826
CB
ILE
A
138
−62.770
45.131
12.238
1.00
30.20
C


ATOM
1828
CG1
ILE
A
138
−62.914
46.576
12.689
1.00
29.92
C


ATOM
1831
CD1
ILE
A
138
−61.681
47.414
12.417
1.00
30.50
C


ATOM
1835
CG2
ILE
A
138
−61.706
44.470
13.085
1.00
29.73
C


ATOM
1839
C
ILE
A
138
−62.140
43.631
10.320
1.00
30.34
C


ATOM
1840
O
ILE
A
138
−60.972
43.203
10.212
1.00
30.65
O


ATOM
1842
N
LEU
A
139
−63.227
42.884
10.098
1.00
29.69
N


ATOM
1843
CA
LEU
A
139
−63.134
41.523
9.562
1.00
29.23
C


ATOM
1845
CB
LEU
A
139
−64.517
40.917
9.332
1.00
28.92
C


ATOM
1848
CG
LEU
A
139
−65.066
40.185
10.545
1.00
29.06
C


ATOM
1850
CD1
LEU
A
139
−66.541
39.875
10.352
1.00
28.20
C


ATOM
1854
CD2
LEU
A
139
−64.250
38.922
10.813
1.00
28.59
C


ATOM
1858
C
LEU
A
139
−62.382
41.546
8.247
1.00
29.00
C


ATOM
1859
O
LEU
A
139
−61.444
40.773
8.035
1.00
29.50
O


ATOM
1861
N
SER
A
140
−62.790
42.458
7.372
1.00
28.14
N


ATOM
1862
CA
SER
A
140
−62.222
42.526
6.054
1.00
27.40
C


ATOM
1864
CB
SER
A
140
−62.912
43.619
5.260
1.00
27.35
C


ATOM
1867
OG
SER
A
140
−63.310
43.103
4.015
1.00
28.43
O


ATOM
1869
C
SER
A
140
−60.720
42.777
6.123
1.00
26.77
C


ATOM
1870
O
SER
A
140
−59.975
42.269
5.298
1.00
27.29
O


ATOM
1872
N
LEU
A
141
−60.285
43.563
7.105
1.00
25.89
N


ATOM
1873
CA
LEU
A
141
−58.872
43.897
7.283
1.00
25.20
C


ATOM
1875
CB
LEU
A
141
−58.717
45.164
8.155
1.00
25.00
C


ATOM
1878
CG
LEU
A
141
−57.298
45.670
8.489
1.00
24.35
C


ATOM
1880
CD1
LEU
A
141
−56.497
45.974
7.232
1.00
22.75
C


ATOM
1884
CD2
LEU
A
141
−57.329
46.893
9.385
1.00
22.81
C


ATOM
1888
C
LEU
A
141
−58.140
42.721
7.920
1.00
24.92
C


ATOM
1889
O
LEU
A
141
−57.034
42.380
7.521
1.00
24.77
O


ATOM
1891
N
TYR
A
142
−58.751
42.101
8.921
1.00
24.51
N


ATOM
1892
CA
TYR
A
142
−58.163
40.913
9.514
1.00
24.36
C


ATOM
1894
CB
TYR
A
142
−59.120
40.325
10.538
1.00
24.18
C


ATOM
1897
CG
TYR
A
142
−58.774
38.941
11.046
1.00
23.72
C


ATOM
1898
CD1
TYR
A
142
−57.878
38.763
12.091
1.00
22.97
C


ATOM
1900
CE1
TYR
A
142
−57.593
37.500
12.582
1.00
24.25
C


ATOM
1902
CZ
TYR
A
142
−58.229
36.386
12.030
1.00
25.75
C


ATOM
1903
OH
TYR
A
142
−57.967
35.100
12.500
1.00
26.97
O


ATOM
1905
CE2
TYR
A
142
−59.120
36.551
10.988
1.00
25.02
C


ATOM
1907
CD2
TYR
A
142
−59.390
37.820
10.511
1.00
24.01
C


ATOM
1909
C
TYR
A
142
−57.877
39.896
8.423
1.00
24.39
C


ATOM
1910
O
TYR
A
142
−56.822
39.276
8.380
1.00
24.19
O


ATOM
1912
N
GLU
A
143
−58.841
39.760
7.527
1.00
24.59
N


ATOM
1913
CA
GLU
A
143
−58.820
38.728
6.521
1.00
24.70
C


ATOM
1915
CB
GLU
A
143
−60.194
38.636
5.853
1.00
24.69
C


ATOM
1918
CG
GLU
A
143
−60.651
37.219
5.547
1.00
27.21
C


ATOM
1921
CD
GLU
A
143
−61.127
36.426
6.774
1.00
30.20
C


ATOM
1922
OE1
GLU
A
143
−60.261
35.776
7.393
1.00
33.40
O


ATOM
1923
OE2
GLU
A
143
−62.351
36.421
7.097
1.00
30.50
O


ATOM
1924
C
GLU
A
143
−57.701
39.010
5.518
1.00
24.08
C


ATOM
1925
O
GLU
A
143
−56.984
38.090
5.126
1.00
24.53
O


ATOM
1927
N
ALA
A
144
−57.537
40.281
5.147
1.00
23.30
N


ATOM
1928
CA
ALA
A
144
−56.515
40.715
4.173
1.00
22.79
C


ATOM
1930
CB
ALA
A
144
−56.787
42.154
3.740
1.00
22.49
C


ATOM
1934
C
ALA
A
144
−55.073
40.598
4.701
1.00
22.42
C


ATOM
1935
O
ALA
A
144
−54.128
40.380
3.935
1.00
22.36
O


ATOM
1937
N
SER
A
145
−54.909
40.713
6.012
1.00
21.80
N


ATOM
1938
CA
SER
A
145
−53.593
40.822
6.600
1.00
21.36
C


ATOM
1940
CB
SER
A
145
−53.726
41.170
8.066
1.00
21.21
C


ATOM
1943
OG
SER
A
145
−54.337
40.100
8.753
1.00
21.59
O


ATOM
1945
C
SER
A
145
−52.797
39.537
6.476
1.00
21.30
C


ATOM
1946
O
SER
A
145
−51.571
39.533
6.701
1.00
21.50
O


ATOM
1948
N
PHE
A
146
−53.488
38.439
6.154
1.00
21.00
N


ATOM
1949
CA
PHE
A
146
−52.827
37.127
6.030
1.00
20.41
C


ATOM
1951
CB
PHE
A
146
−53.796
35.979
6.338
1.00
20.20
C


ATOM
1954
CG
PHE
A
146
−54.130
35.873
7.798
1.00
19.91
C


ATOM
1955
CD1
PHE
A
146
−53.382
35.087
8.637
1.00
20.48
C


ATOM
1957
CE1
PHE
A
146
−53.679
35.012
9.992
1.00
20.59
C


ATOM
1959
CZ
PHE
A
146
−54.731
35.740
10.508
1.00
19.15
C


ATOM
1961
CE2
PHE
A
146
−55.464
36.536
9.691
1.00
19.10
C


ATOM
1963
CD2
PHE
A
146
−55.155
36.615
8.343
1.00
20.16
C


ATOM
1965
C
PHE
A
146
−52.174
36.964
4.680
1.00
19.76
C


ATOM
1966
O
PHE
A
146
−51.305
36.116
4.523
1.00
19.46
O


ATOM
1968
N
LEU
A
147
−52.550
37.819
3.729
1.00
19.34
N


ATOM
1969
CA
LEU
A
147
−51.933
37.804
2.402
1.00
19.13
C


ATOM
1971
CB
LEU
A
147
−52.905
38.367
1.347
1.00
18.67
C


ATOM
1974
CG
LEU
A
147
−53.964
37.334
.919
1.00
18.11
C


ATOM
1976
CD1
LEU
A
147
−55.090
37.248
1.961
1.00
15.10
C


ATOM
1980
CD2
LEU
A
147
−54.494
37.609
−.499
1.00
16.50
C


ATOM
1984
C
LEU
A
147
−50.582
38.525
2.369
1.00
19.10
C


ATOM
1985
O
LEU
A
147
−49.992
38.715
1.311
1.00
19.44
O


ATOM
1987
N
ALA
A
148
−50.075
38.882
3.542
1.00
19.13
N


ATOM
1988
CA
ALA
A
148
−48.967
39.806
3.664
1.00
18.84
C


ATOM
1990
CB
ALA
A
148
−48.762
40.167
5.117
1.00
18.78
C


ATOM
1994
C
ALA
A
148
−47.723
39.184
3.113
1.00
18.85
C


ATOM
1995
O
ALA
A
148
−47.548
37.975
3.206
1.00
18.80
O


ATOM
1997
N
LEU
A
149
−46.869
40.013
2.523
1.00
19.08
N


ATOM
1998
CA
LEU
A
149
−45.507
39.601
2.215
1.00
19.32
C


ATOM
2000
CB
LEU
A
149
−45.056
40.145
.863
1.00
19.14
C


ATOM
2003
CG
LEU
A
149
−45.819
39.650
−.364
1.00
19.16
C


ATOM
2005
CD1
LEU
A
149
−45.036
39.929
−1.645
1.00
18.43
C


ATOM
2009
CD2
LEU
A
149
−46.089
38.175
−.259
1.00
20.09
C


ATOM
2013
C
LEU
A
149
−44.527
40.029
3.304
1.00
19.60
C


ATOM
2014
O
LEU
A
149
−44.797
40.916
4.113
1.00
19.20
O


ATOM
2016
N
GLU
A
150
−43.384
39.361
3.309
1.00
20.42
N


ATOM
2017
CA
GLU
A
150
−42.286
39.674
4.215
1.00
21.03
C


ATOM
2019
CB
GLU
A
150
−41.098
38.741
3.948
1.00
21.03
C


ATOM
2022
CG
GLU
A
150
−40.491
38.176
5.205
1.00
22.23
C


ATOM
2025
CD
GLU
A
150
−39.433
37.117
4.941
1.00
24.34
C


ATOM
2026
OE1
GLU
A
150
−39.189
36.786
3.753
1.00
24.53
O


ATOM
2027
OE2
GLU
A
150
−38.843
36.625
5.940
1.00
25.82
O


ATOM
2028
C
GLU
A
150
−41.887
41.130
4.011
1.00
21.29
C


ATOM
2029
O
GLU
A
150
−41.619
41.544
2.885
1.00
21.13
O


ATOM
2031
N
GLY
A
151
−41.893
41.903
5.094
1.00
21.89
N


ATOM
2032
CA
GLY
A
151
−41.636
43.341
5.030
1.00
22.25
C


ATOM
2035
C
GLY
A
151
−42.871
44.221
5.152
1.00
22.67
C


ATOM
2036
O
GLY
A
151
−42.757
45.397
5.462
1.00
22.84
O


ATOM
2038
N
GLU
A
152
−44.055
43.669
4.918
1.00
23.19
N


ATOM
2039
CA
GLU
A
152
−45.263
44.474
4.937
1.00
23.76
C


ATOM
2041
CB
GLU
A
152
−46.350
43.813
4.100
1.00
23.79
C


ATOM
2044
CG
GLU
A
152
−46.037
43.890
2.615
1.00
23.98
C


ATOM
2047
CD
GLU
A
152
−47.147
43.368
1.749
1.00
23.72
C


ATOM
2048
OE1
GLU
A
152
−47.226
43.771
.574
1.00
22.11
O


ATOM
2049
OE2
GLU
A
152
−47.939
42.542
2.241
1.00
25.41
O


ATOM
2050
C
GLU
A
152
−45.722
44.719
6.365
1.00
24.47
C


ATOM
2051
O
GLU
A
152
−46.586
44.006
6.906
1.00
24.70
O


ATOM
2053
N
ASN
A
153
−45.134
45.744
6.969
1.00
25.00
N


ATOM
2054
CA
ASN
A
153
−45.287
45.978
8.404
1.00
25.59
C


ATOM
2056
CB
ASN
A
153
−44.221
46.957
8.902
1.00
25.55
C


ATOM
2059
CG
ASN
A
153
−44.386
48.331
8.308
1.00
25.57
C


ATOM
2060
OD1
ASN
A
153
−44.114
48.557
7.128
1.00
24.52
O


ATOM
2061
ND2
ASN
A
153
−44.867
49.256
9.117
1.00
27.05
N


ATOM
2064
C
ASN
A
153
−46.679
46.486
8.772
1.00
25.92
C


ATOM
2065
O
ASN
A
153
−47.172
46.238
9.872
1.00
26.02
O


ATOM
2067
N
ILE
A
154
−47.310
47.197
7.847
1.00
26.35
N


ATOM
2068
CA
ILE
A
154
−48.638
47.751
8.082
1.00
26.57
C


ATOM
2070
CB
ILE
A
154
−49.013
48.779
7.007
1.00
26.45
C


ATOM
2072
CG1
ILE
A
154
−48.159
50.023
7.164
1.00
26.36
C


ATOM
2075
CD1
ILE
A
154
−48.060
50.790
5.883
1.00
27.92
C


ATOM
2079
CG2
ILE
A
154
−50.467
49.152
7.089
1.00
26.25
C


ATOM
2083
C
ILE
A
154
−49.674
46.643
8.111
1.00
26.96
C


ATOM
2084
O
ILE
A
154
−50.650
46.741
8.851
1.00
27.12
O


ATOM
2086
N
LEU
A
155
−49.467
45.588
7.320
1.00
27.35
N


ATOM
2087
CA
LEU
A
155
−50.422
44.475
7.304
1.00
27.76
C


ATOM
2089
CB
LEU
A
155
−50.238
43.594
6.067
1.00
27.73
C


ATOM
2092
CG
LEU
A
155
−51.090
43.990
4.875
1.00
27.31
C


ATOM
2094
CD1
LEU
A
155
−50.980
45.456
4.683
1.00
28.13
C


ATOM
2098
CD2
LEU
A
155
−50.633
43.262
3.623
1.00
27.79
C


ATOM
2102
C
LEU
A
155
−50.340
43.642
8.582
1.00
28.22
C


ATOM
2103
O
LEU
A
155
−51.361
43.237
9.112
1.00
27.95
O


ATOM
2105
N
ASP
A
156
−49.128
43.395
9.068
1.00
28.95
N


ATOM
2106
CA
ASP
A
156
−48.944
42.723
10.348
1.00
29.71
C


ATOM
2108
CB
ASP
A
156
−47.450
42.461
10.639
1.00
30.23
C


ATOM
2111
CG
ASP
A
156
−46.938
41.125
10.027
1.00
32.82
C


ATOM
2112
OD1
ASP
A
156
−47.765
40.240
9.658
1.00
36.27
O


ATOM
2113
OD2
ASP
A
156
−45.700
40.961
9.923
1.00
34.48
O


ATOM
2114
C
ASP
A
156
−49.570
43.539
11.469
1.00
29.66
C


ATOM
2115
O
ASP
A
156
−50.109
42.969
12.425
1.00
29.81
O


ATOM
2117
N
GLU
A
157
−49.514
44.866
11.345
1.00
29.69
N


ATOM
2118
CA
GLU
A
157
−50.129
45.768
12.329
1.00
29.57
C


ATOM
2120
CB
GLU
A
157
−49.581
47.186
12.178
1.00
29.71
C


ATOM
2123
CG
GLU
A
157
−48.164
47.326
12.712
1.00
30.53
C


ATOM
2126
CD
GLU
A
157
−47.455
48.544
12.169
1.00
31.96
C


ATOM
2127
OE1
GLU
A
157
−48.167
49.491
11.741
1.00
33.56
O


ATOM
2128
OE2
GLU
A
157
−46.195
48.546
12.166
1.00
31.54
O


ATOM
2129
C
GLU
A
157
−51.649
45.771
12.230
1.00
29.16
C


ATOM
2130
O
GLU
A
157
−52.330
45.819
13.241
1.00
29.00
O


ATOM
2132
N
ALA
A
158
−52.172
45.716
11.010
1.00
28.86
N


ATOM
2133
CA
ALA
A
158
−53.606
45.570
10.792
1.00
28.79
C


ATOM
2135
CB
ALA
A
158
−53.893
45.456
9.301
1.00
28.44
C


ATOM
2139
C
ALA
A
158
−54.171
44.352
11.536
1.00
28.93
C


ATOM
2140
O
ALA
A
158
−55.265
44.402
12.098
1.00
28.77
O


ATOM
2142
N
LYS
A
159
−53.416
43.260
11.528
1.00
29.19
N


ATOM
2143
CA
LYS
A
159
−53.849
42.022
12.143
1.00
29.57
C


ATOM
2145
CB
LYS
A
159
−52.929
40.880
11.710
1.00
29.62
C


ATOM
2148
CG
LYS
A
159
−53.297
39.513
12.258
1.00
30.27
C


ATOM
2151
CD
LYS
A
159
−52.765
38.365
11.387
1.00
31.78
C


ATOM
2154
CE
LYS
A
159
−51.259
38.132
11.525
1.00
33.15
C


ATOM
2157
NZ
LYS
A
159
−50.911
36.696
11.248
1.00
33.88
N


ATOM
2161
C
LYS
A
159
−53.866
42.160
13.670
1.00
29.99
C


ATOM
2162
O
LYS
A
159
−54.767
41.642
14.332
1.00
30.29
O


ATOM
2164
N
VAL
A
160
−52.886
42.862
14.236
1.00
30.07
N


ATOM
2165
CA
VAL
A
160
−52.840
43.047
15.696
1.00
30.03
C


ATOM
2167
CB
VAL
A
160
−51.520
43.691
16.162
1.00
29.88
C


ATOM
2169
CG1
VAL
A
160
−51.515
43.863
17.671
1.00
28.94
C


ATOM
2173
CG2
VAL
A
160
−50.341
42.848
15.707
1.00
30.23
C


ATOM
2177
C
VAL
A
160
−53.996
43.925
16.149
1.00
30.05
C


ATOM
2178
O
VAL
A
160
−54.544
43.752
17.234
1.00
30.31
O


ATOM
2180
N
PHE
A
161
−54.355
44.862
15.290
1.00
30.08
N


ATOM
2181
CA
PHE
A
161
−55.407
45.808
15.558
1.00
30.09
C


ATOM
2183
CB
PHE
A
161
−55.255
46.993
14.598
1.00
29.92
C


ATOM
2186
CG
PHE
A
161
−56.370
47.974
14.654
1.00
28.52
C


ATOM
2187
CD1
PHE
A
161
−56.347
49.005
15.554
1.00
27.92
C


ATOM
2189
CE1
PHE
A
161
−57.361
49.903
15.600
1.00
28.06
C


ATOM
2191
CZ
PHE
A
161
−58.411
49.783
14.733
1.00
28.76
C


ATOM
2193
CE2
PHE
A
161
−58.441
48.757
13.826
1.00
28.31
C


ATOM
2195
CD2
PHE
A
161
−57.429
47.869
13.789
1.00
28.16
C


ATOM
2197
C
PHE
A
161
−56.746
45.123
15.388
1.00
30.55
C


ATOM
2198
O
PHE
A
161
−57.598
45.203
16.258
1.00
30.53
O


ATOM
2200
N
ALA
A
162
−56.930
44.438
14.271
1.00
31.34
N


ATOM
2201
CA
ALA
A
162
−58.193
43.770
14.023
1.00
32.26
C


ATOM
2203
CB
ALA
A
162
−58.217
43.139
12.643
1.00
32.31
C


ATOM
2207
C
ALA
A
162
−58.500
42.734
15.118
1.00
33.09
C


ATOM
2208
O
ALA
A
162
−59.557
42.814
15.735
1.00
33.83
O


ATOM
2210
N
ILE
A
163
−57.587
41.799
15.393
1.00
33.59
N


ATOM
2211
CA
ILE
A
163
−57.826
40.799
16.438
1.00
34.02
C


ATOM
2213
CB
ILE
A
163
−56.597
39.907
16.745
1.00
33.98
C


ATOM
2215
CG1
ILE
A
163
−56.235
39.008
15.566
1.00
34.37
C


ATOM
2218
CD1
ILE
A
163
−54.878
38.317
15.722
1.00
34.38
C


ATOM
2222
CG2
ILE
A
163
−56.883
38.995
17.919
1.00
33.38
C


ATOM
2226
C
ILE
A
163
−58.225
41.472
17.744
1.00
34.75
C


ATOM
2227
O
ILE
A
163
−59.189
41.064
18.373
1.00
35.03
O


ATOM
2229
N
SER
A
164
−57.501
42.504
18.156
1.00
35.76
N


ATOM
2230
CA
SER
A
164
−57.763
43.117
19.463
1.00
36.59
C


ATOM
2232
CB
SER
A
164
−56.828
44.293
19.735
1.00
36.59
C


ATOM
2235
OG
SER
A
164
−57.350
45.485
19.164
1.00
36.46
O


ATOM
2237
C
SER
A
164
−59.196
43.611
19.556
1.00
37.42
C


ATOM
2238
O
SER
A
164
−59.860
43.420
20.571
1.00
37.38
O


ATOM
2240
N
HIS
A
165
−59.661
44.251
18.486
1.00
38.51
N


ATOM
2241
CA
HIS
A
165
−60.990
44.851
18.467
1.00
39.40
C


ATOM
2243
CB
HIS
A
165
−61.008
46.097
17.574
1.00
39.76
C


ATOM
2246
CG
HIS
A
165
−60.467
47.323
18.251
1.00
42.06
C


ATOM
2247
ND1
HIS
A
165
−59.205
47.821
18.000
1.00
44.21
N


ATOM
2249
CE1
HIS
A
165
−58.998
48.893
18.748
1.00
45.04
C


ATOM
2251
NE2
HIS
A
165
−60.080
49.106
19.478
1.00
44.99
N


ATOM
2253
CD2
HIS
A
165
−61.011
48.135
19.192
1.00
43.76
C


ATOM
2255
C
HIS
A
165
−62.078
43.872
18.060
1.00
39.38
C


ATOM
2256
O
HIS
A
165
−63.248
44.244
18.067
1.00
39.43
O


ATOM
2258
N
LEU
A
166
−61.683
42.632
17.745
1.00
39.54
N


ATOM
2259
CA
LEU
A
166
−62.589
41.550
17.326
1.00
39.55
C


ATOM
2261
CB
LEU
A
166
−62.001
40.766
16.154
1.00
39.28
C


ATOM
2264
CG
LEU
A
166
−62.156
41.339
14.752
1.00
38.23
C


ATOM
2266
CD1
LEU
A
166
−61.361
40.501
13.805
1.00
37.53
C


ATOM
2270
CD2
LEU
A
166
−63.594
41.367
14.332
1.00
37.21
C


ATOM
2274
C
LEU
A
166
−62.842
40.555
18.438
1.00
40.08
C


ATOM
2275
O
LEU
A
166
−63.991
40.220
18.733
1.00
40.42
O


ATOM
2277
N
LYS
A
167
−61.766
40.074
19.052
1.00
40.58
N


ATOM
2278
CA
LYS
A
167
−61.875
39.090
20.133
1.00
41.09
C


ATOM
2280
CB
LYS
A
167
−60.491
38.632
20.595
1.00
41.13
C


ATOM
2283
CG
LYS
A
167
−59.897
39.399
21.768
1.00
42.16
C


ATOM
2286
CD
LYS
A
167
−58.421
39.035
22.013
1.00
44.45
C


ATOM
2289
CE
LYS
A
167
−58.070
37.561
21.654
1.00
45.56
C


ATOM
2292
NZ
LYS
A
167
−56.610
37.237
21.798
1.00
46.20
N


ATOM
2296
C
LYS
A
167
−62.678
39.584
21.337
1.00
41.23
C


ATOM
2297
O
LYS
A
167
−62.994
38.793
22.215
1.00
41.47
O


ATOM
2299
N
GLU
A
168
−62.995
40.884
21.352
1.00
41.49
N


ATOM
2300
CA
GLU
A
168
−63.704
41.583
22.437
1.00
41.42
C


ATOM
2302
CB
GLU
A
168
−65.201
41.730
22.100
1.00
41.27
C


ATOM
2305
CG
GLU
A
168
−65.995
40.424
22.016
1.00
41.01
C


ATOM
2308
CD
GLU
A
168
−66.928
40.328
20.787
1.00
40.84
C


ATOM
2309
OE1
GLU
A
168
−66.701
41.040
19.770
1.00
39.82
O


ATOM
2310
OE2
GLU
A
168
−67.884
39.509
20.845
1.00
39.04
O


ATOM
2311
C
GLU
A
168
−63.484
41.004
23.843
1.00
41.66
C


ATOM
2312
O
GLU
A
168
−62.435
41.231
24.469
1.00
41.46
O


ATOM
2314
N
GLU
A
172
−71.668
38.961
21.192
1.00
55.65
N


ATOM
2315
CA
GLU
A
172
−73.052
39.153
20.721
1.00
56.00
C


ATOM
2317
CB
GLU
A
172
−73.876
37.880
20.937
1.00
56.27
C


ATOM
2320
CG
GLU
A
172
−73.198
36.620
20.390
1.00
57.12
C


ATOM
2323
CD
GLU
A
172
−74.056
35.363
20.516
1.00
58.18
C


ATOM
2324
OE1
GLU
A
172
−75.300
35.480
20.622
1.00
58.84
O


ATOM
2325
OE2
GLU
A
172
−73.480
34.249
20.498
1.00
58.98
O


ATOM
2326
C
GLU
A
172
−73.766
40.386
21.334
1.00
55.70
C


ATOM
2327
O
GLU
A
172
−74.956
40.347
21.688
1.00
54.95
O


ATOM
2329
N
LYS
A
173
−72.987
41.458
21.486
1.00
55.58
N


ATOM
2330
CA
LYS
A
173
−73.496
42.831
21.466
1.00
55.43
C


ATOM
2332
CB
LYS
A
173
−72.649
43.741
22.357
1.00
55.51
C


ATOM
2335
CG
LYS
A
173
−72.527
43.223
23.793
1.00
56.07
C


ATOM
2338
CD
LYS
A
173
−72.803
44.291
24.868
1.00
55.87
C


ATOM
2341
CE
LYS
A
173
−73.111
43.642
26.221
1.00
55.20
C


ATOM
2344
NZ
LYS
A
173
−72.893
44.576
27.348
1.00
54.63
N


ATOM
2348
C
LYS
A
173
−73.449
43.297
20.009
1.00
55.01
C


ATOM
2349
O
LYS
A
173
−74.091
44.274
19.625
1.00
54.97
O


ATOM
2351
N
ILE
A
174
−72.647
42.581
19.223
1.00
54.57
N


ATOM
2352
CA
ILE
A
174
−72.713
42.550
17.767
1.00
54.22
C


ATOM
2354
CB
ILE
A
174
−71.429
41.845
17.227
1.00
54.21
C


ATOM
2356
CG1
ILE
A
174
−70.196
42.724
17.474
1.00
54.73
C


ATOM
2359
CD1
ILE
A
174
−68.858
41.949
17.553
1.00
55.78
C


ATOM
2363
CG2
ILE
A
174
−71.532
41.504
15.759
1.00
54.06
C


ATOM
2367
C
ILE
A
174
−73.957
41.744
17.384
1.00
53.88
C


ATOM
2368
O
ILE
A
174
−74.587
41.134
18.250
1.00
53.83
O


ATOM
2370
N
GLY
A
175
−74.332
41.747
16.107
1.00
53.59
N


ATOM
2371
CA
GLY
A
175
−75.351
40.817
15.613
1.00
53.51
C


ATOM
2374
C
GLY
A
175
−75.130
39.395
16.126
1.00
53.43
C


ATOM
2375
O
GLY
A
175
−74.119
39.105
16.760
1.00
53.80
O


ATOM
2377
N
LYS
A
176
−76.070
38.495
15.861
1.00
53.10
N


ATOM
2378
CA
LYS
A
176
−75.926
37.098
16.286
1.00
52.68
C


ATOM
2380
CB
LYS
A
176
−77.291
36.522
16.677
1.00
52.99
C


ATOM
2383
CG
LYS
A
176
−78.003
37.387
17.746
1.00
54.12
C


ATOM
2386
CD
LYS
A
176
−79.201
36.697
18.436
1.00
54.57
C


ATOM
2389
CE
LYS
A
176
−79.503
37.369
19.792
1.00
54.79
C


ATOM
2392
NZ
LYS
A
176
−80.458
36.598
20.653
1.00
55.20
N


ATOM
2396
C
LYS
A
176
−75.246
36.267
15.196
1.00
51.83
C


ATOM
2397
O
LYS
A
176
−74.393
35.433
15.486
1.00
51.69
O


ATOM
2399
N
GLU
A
177
−75.609
36.520
13.940
1.00
50.90
N


ATOM
2400
CA
GLU
A
177
−74.975
35.854
12.800
1.00
50.19
C


ATOM
2402
CB
GLU
A
177
−75.792
36.062
11.507
1.00
50.41
C


ATOM
2405
CG
GLU
A
177
−75.506
37.368
10.735
1.00
51.03
C


ATOM
2408
CD
GLU
A
177
−76.403
37.572
9.507
1.00
52.17
C


ATOM
2409
OE1
GLU
A
177
−75.955
38.276
8.569
1.00
52.63
O


ATOM
2410
OE2
GLU
A
177
−77.546
37.048
9.479
1.00
52.03
O


ATOM
2411
C
GLU
A
177
−73.549
36.369
12.607
1.00
49.10
C


ATOM
2412
O
GLU
A
177
−72.626
35.597
12.321
1.00
49.53
O


ATOM
2414
N
LEU
A
178
−73.388
37.681
12.770
1.00
47.39
N


ATOM
2415
CA
LEU
A
178
−72.110
38.371
12.575
1.00
45.63
C


ATOM
2417
CB
LEU
A
178
−72.352
39.877
12.615
1.00
45.59
C


ATOM
2420
CG
LEU
A
178
−71.280
40.809
12.085
1.00
45.45
C


ATOM
2422
CD1
LEU
A
178
−70.969
40.515
10.620
1.00
45.45
C


ATOM
2426
CD2
LEU
A
178
−71.765
42.233
12.279
1.00
44.63
C


ATOM
2430
C
LEU
A
178
−71.095
37.979
13.634
1.00
44.06
C


ATOM
2431
O
LEU
A
178
−69.902
38.012
13.387
1.00
43.70
O


ATOM
2433
N
ALA
A
179
−71.586
37.611
14.810
1.00
42.54
N


ATOM
2434
CA
ALA
A
179
−70.753
37.041
15.847
1.00
41.66
C


ATOM
2436
CB
ALA
A
179
−71.545
36.884
17.122
1.00
41.69
C


ATOM
2440
C
ALA
A
179
−70.193
35.695
15.416
1.00
40.75
C


ATOM
2441
O
ALA
A
179
−69.078
35.351
15.769
1.00
40.61
O


ATOM
2443
N
GLU
A
180
−70.974
34.931
14.664
1.00
39.88
N


ATOM
2444
CA
GLU
A
180
−70.518
33.639
14.143
1.00
39.40
C


ATOM
2446
CB
GLU
A
180
−71.710
32.781
13.718
1.00
39.74
C


ATOM
2449
CG
GLU
A
180
−72.650
32.415
14.865
1.00
40.88
C


ATOM
2452
CD
GLU
A
180
−73.913
31.710
14.398
1.00
42.02
C


ATOM
2453
OE1
GLU
A
180
−74.380
32.022
13.271
1.00
43.05
O


ATOM
2454
OE2
GLU
A
180
−74.437
30.857
15.166
1.00
41.93
O


ATOM
2455
C
GLU
A
180
−69.565
33.805
12.960
1.00
38.29
C


ATOM
2456
O
GLU
A
180
−68.739
32.940
12.699
1.00
38.23
O


ATOM
2458
N
GLN
A
181
−69.704
34.911
12.240
1.00
36.92
N


ATOM
2459
CA
GLN
A
181
−68.748
35.299
11.223
1.00
35.74
C


ATOM
2461
CB
GLN
A
181
−69.272
36.516
10.463
1.00
35.94
C


ATOM
2464
CG
GLN
A
181
−68.931
36.537
8.975
1.00
36.93
C


ATOM
2467
CD
GLN
A
181
−69.693
35.492
8.176
1.00
37.72
C


ATOM
2468
OE1
GLN
A
181
−70.769
35.067
8.567
1.00
38.78
O


ATOM
2469
NE2
GLN
A
181
−69.139
35.083
7.050
1.00
38.11
N


ATOM
2472
C
GLN
A
181
−67.406
35.632
11.881
1.00
34.64
C


ATOM
2473
O
GLN
A
181
−66.348
35.411
11.294
1.00
34.73
O


ATOM
2475
N
VAL
A
182
−67.448
36.158
13.102
1.00
33.29
N


ATOM
2476
CA
VAL
A
182
−66.224
36.488
13.842
1.00
32.28
C


ATOM
2478
CB
VAL
A
182
−66.433
37.653
14.855
1.00
32.10
C


ATOM
2480
CG1
VAL
A
182
−66.328
38.991
14.166
1.00
31.51
C


ATOM
2484
CG2
VAL
A
182
−65.420
37.594
15.957
1.00
31.69
C


ATOM
2488
C
VAL
A
182
−65.613
35.298
14.575
1.00
31.49
C


ATOM
2489
O
VAL
A
182
−64.396
35.230
14.668
1.00
31.43
O


ATOM
2491
N
SER
A
183
−66.422
34.378
15.106
1.00
30.56
N


ATOM
2492
CA
SER
A
183
−65.860
33.174
15.744
1.00
30.16
C


ATOM
2494
CB
SER
A
183
−66.926
32.253
16.327
1.00
30.29
C


ATOM
2497
OG
SER
A
183
−67.796
32.934
17.214
1.00
32.35
O


ATOM
2499
C
SER
A
183
−65.111
32.403
14.692
1.00
29.30
C


ATOM
2500
O
SER
A
183
−63.974
31.981
14.903
1.00
29.32
O


ATOM
2502
N
HIS
A
184
−65.768
32.252
13.546
1.00
28.24
N


ATOM
2503
CA
HIS
A
184
−65.215
31.578
12.379
1.00
27.36
C


ATOM
2505
CB
HIS
A
184
−66.245
31.653
11.254
1.00
27.41
C


ATOM
2508
CG
HIS
A
184
−65.930
30.807
10.064
1.00
26.90
C


ATOM
2509
ND1
HIS
A
184
−65.484
29.511
10.166
1.00
27.76
N


ATOM
2511
CE1
HIS
A
184
−65.321
29.009
8.955
1.00
27.47
C


ATOM
2513
NE2
HIS
A
184
−65.661
29.929
8.075
1.00
26.52
N


ATOM
2515
CD2
HIS
A
184
−66.059
31.058
8.742
1.00
26.61
C


ATOM
2517
C
HIS
A
184
−63.898
32.200
11.940
1.00
26.69
C


ATOM
2518
O
HIS
A
184
−62.881
31.525
11.862
1.00
26.44
O


ATOM
2520
N
ALA
A
185
−63.909
33.498
11.675
1.00
26.11
N


ATOM
2521
CA
ALA
A
185
−62.711
34.176
11.205
1.00
25.63
C


ATOM
2523
CB
ALA
A
185
−62.970
35.627
11.109
1.00
25.74
C


ATOM
2527
C
ALA
A
185
−61.552
33.917
12.153
1.00
25.33
C


ATOM
2528
O
ALA
A
185
−60.451
33.572
11.725
1.00
25.79
O


ATOM
2530
N
LEU
A
186
−61.814
34.066
13.450
1.00
24.77
N


ATOM
2531
CA
LEU
A
186
−60.776
33.940
14.470
1.00
24.05
C


ATOM
2533
CB
LEU
A
186
−61.256
34.498
15.814
1.00
23.63
C


ATOM
2536
CG
LEU
A
186
−61.516
36.012
15.903
1.00
22.51
C


ATOM
2538
CD1
LEU
A
186
−62.057
36.399
17.266
1.00
20.79
C


ATOM
2542
CD2
LEU
A
186
−60.273
36.793
15.613
1.00
21.13
C


ATOM
2546
C
LEU
A
186
−60.316
32.497
14.613
1.00
24.02
C


ATOM
2547
O
LEU
A
186
−59.169
32.250
14.943
1.00
24.27
O


ATOM
2549
N
GLU
A
187
−61.201
31.543
14.347
1.00
23.88
N


ATOM
2550
CA
GLU
A
187
−60.836
30.117
14.348
1.00
23.53
C


ATOM
2552
CB
GLU
A
187
−62.029
29.270
13.919
1.00
23.66
C


ATOM
2555
CG
GLU
A
187
−61.945
27.828
14.316
1.00
25.26
C


ATOM
2558
CD
GLU
A
187
−63.049
26.971
13.693
1.00
28.04
C


ATOM
2559
OE1
GLU
A
187
−63.672
27.393
12.681
1.00
28.67
O


ATOM
2560
OE2
GLU
A
187
−63.288
25.858
14.225
1.00
29.72
O


ATOM
2561
C
GLU
A
187
−59.686
29.838
13.395
1.00
22.92
C


ATOM
2562
O
GLU
A
187
−58.765
29.091
13.729
1.00
22.53
O


ATOM
2564
N
LEU
A
188
−59.763
30.446
12.209
1.00
22.46
N


ATOM
2565
CA
LEU
A
188
−58.799
30.246
11.121
1.00
22.07
C


ATOM
2567
CB
LEU
A
188
−58.802
28.794
10.635
1.00
22.11
C


ATOM
2570
CG
LEU
A
188
−57.876
28.362
9.493
1.00
22.17
C


ATOM
2572
CD1
LEU
A
188
−56.456
28.051
9.961
1.00
21.62
C


ATOM
2576
CD2
LEU
A
188
−58.464
27.134
8.822
1.00
22.40
C


ATOM
2580
C
LEU
A
188
−59.237
31.137
9.985
1.00
21.69
C


ATOM
2581
O
LEU
A
188
−60.380
31.075
9.563
1.00
21.53
O


ATOM
2583
N
PRO
A
189
−58.333
31.973
9.474
1.00
21.46
N


ATOM
2584
CA
PRO
A
189
−58.745
32.935
8.472
1.00
21.18
C


ATOM
2586
CB
PRO
A
189
−57.592
33.936
8.486
1.00
21.03
C


ATOM
2589
CG
PRO
A
189
−56.423
33.118
8.757
1.00
21.21
C


ATOM
2592
CD
PRO
A
189
−56.873
31.992
9.668
1.00
21.61
C


ATOM
2595
C
PRO
A
189
−58.946
32.314
7.076
1.00
20.96
C


ATOM
2596
O
PRO
A
189
−58.418
31.247
6.793
1.00
21.02
O


ATOM
2597
N
LEU
A
190
−59.714
32.994
6.226
1.00
20.89
N


ATOM
2598
CA
LEU
A
190
−60.018
32.533
4.867
1.00
20.74
C


ATOM
2600
CB
LEU
A
190
−60.597
33.690
4.053
1.00
20.76
C


ATOM
2603
CG
LEU
A
190
−62.090
33.924
4.259
1.00
21.35
C


ATOM
2605
CD1
LEU
A
190
−62.537
35.196
3.578
1.00
20.54
C


ATOM
2609
CD2
LEU
A
190
−62.889
32.712
3.720
1.00
23.39
C


ATOM
2613
C
LEU
A
190
−58.814
31.971
4.112
1.00
20.35
C


ATOM
2614
O
LEU
A
190
−58.859
30.871
3.546
1.00
20.08
O


ATOM
2616
N
HIS
A
191
−57.734
32.746
4.132
1.00
19.81
N


ATOM
2617
CA
HIS
A
191
−56.537
32.448
3.364
1.00
18.89
C


ATOM
2619
CB
HIS
A
191
−55.548
33.592
3.536
1.00
18.92
C


ATOM
2622
CG
HIS
A
191
−54.372
33.520
2.623
1.00
18.51
C


ATOM
2623
ND1
HIS
A
191
−54.482
33.647
1.258
1.00
18.08
N


ATOM
2625
CE1
HIS
A
191
−53.281
33.544
.718
1.00
18.96
C


ATOM
2627
NE2
HIS
A
191
−52.397
33.371
1.684
1.00
17.01
N


ATOM
2629
CD2
HIS
A
191
−53.054
33.355
2.884
1.00
17.94
C


ATOM
2631
C
HIS
A
191
−55.889
31.142
3.759
1.00
18.32
C


ATOM
2632
O
HIS
A
191
−55.217
30.531
2.942
1.00
17.98
O


ATOM
2634
N
ARG
A
192
−56.093
30.719
5.006
1.00
18.06
N


ATOM
2635
CA
ARG
A
192
−55.502
29.480
5.523
1.00
17.99
C


ATOM
2637
CB
ARG
A
192
−54.940
29.722
6.909
1.00
17.77
C


ATOM
2640
CG
ARG
A
192
−53.822
30.683
6.881
1.00
18.47
C


ATOM
2643
CD
ARG
A
192
−53.082
30.748
8.196
1.00
19.80
C


ATOM
2646
NE
ARG
A
192
−52.174
31.900
8.224
1.00
20.33
N


ATOM
2648
CZ
ARG
A
192
−51.176
32.048
9.077
1.00
20.10
C


ATOM
2649
NH1
ARG
A
192
−50.928
31.113
9.972
1.00
21.72
N


ATOM
2652
NH2
ARG
A
192
−50.407
33.119
9.019
1.00
20.79
N


ATOM
2655
C
ARG
A
192
−56.477
28.311
5.590
1.00
18.00
C


ATOM
2656
O
ARG
A
192
−56.067
27.170
5.871
1.00
18.21
O


ATOM
2658
N
ARG
A
193
−57.758
28.588
5.335
1.00
17.59
N


ATOM
2659
CA
ARG
A
193
−58.779
27.563
5.382
1.00
17.16
C


ATOM
2661
CB
ARG
A
193
−60.141
28.164
5.741
1.00
17.45
C


ATOM
2664
CG
ARG
A
193
−61.109
27.132
6.322
1.00
18.70
C


ATOM
2667
CD
ARG
A
193
−62.479
27.682
6.686
1.00
20.20
C


ATOM
2670
NE
ARG
A
193
−62.423
28.682
7.751
1.00
22.59
N


ATOM
2672
CZ
ARG
A
193
−62.386
28.435
9.067
1.00
24.12
C


ATOM
2673
NH1
ARG
A
193
−62.395
27.198
9.575
1.00
22.66
N


ATOM
2676
NH2
ARG
A
193
−62.348
29.469
9.898
1.00
26.36
N


ATOM
2679
C
ARG
A
193
−58.871
26.842
4.058
1.00
16.63
C


ATOM
2680
O
ARG
A
193
−58.861
27.461
3.005
1.00
15.93
O


ATOM
2682
N
THR
A
194
−58.955
25.518
4.149
1.00
16.72
N


ATOM
2683
CA
THR
A
194
−59.332
24.638
3.051
1.00
16.88
C


ATOM
2685
CB
THR
A
194
−59.520
23.250
3.592
1.00
16.43
C


ATOM
2687
OG1
THR
A
194
−58.252
22.605
3.583
1.00
17.74
O


ATOM
2689
CG2
THR
A
194
−60.454
22.441
2.754
1.00
18.43
C


ATOM
2693
C
THR
A
194
−60.607
25.112
2.384
1.00
17.28
C


ATOM
2694
O
THR
A
194
−61.441
25.734
3.022
1.00
17.44
O


ATOM
2696
N
GLN
A
195
−60.765
24.855
1.091
1.00
18.05
N


ATOM
2697
CA
GLN
A
195
−61.925
25.402
.397
1.00
18.76
C


ATOM
2699
CB
GLN
A
195
−61.768
25.381
−1.114
1.00
18.87
C


ATOM
2702
CG
GLN
A
195
−63.145
25.523
−1.761
1.00
20.75
C


ATOM
2705
CD
GLN
A
195
−63.095
25.922
−3.179
1.00
23.41
C


ATOM
2706
OE1
GLN
A
195
−62.031
26.088
−3.733
1.00
27.07
O


ATOM
2707
NE2
GLN
A
195
−64.248
26.090
−3.791
1.00
25.02
N


ATOM
2710
C
GLN
A
195
−63.243
24.704
.765
1.00
18.93
C


ATOM
2711
O
GLN
A
195
−64.169
25.333
1.291
1.00
19.42
O


ATOM
2713
N
ARG
A
196
−63.345
23.418
.443
1.00
18.90
N


ATOM
2714
CA
ARG
A
196
−64.577
22.685
.663
1.00
18.68
C


ATOM
2716
CB
ARG
A
196
−64.356
21.186
.454
1.00
18.53
C


ATOM
2719
CG
ARG
A
196
−64.851
20.644
−.888
1.00
18.31
C


ATOM
2722
CD
ARG
A
196
−64.705
21.622
−2.037
1.00
17.93
C


ATOM
2725
NE
ARG
A
196
−65.930
21.720
−2.834
1.00
18.52
N


ATOM
2727
CZ
ARG
A
196
−66.480
22.857
−3.256
1.00
18.63
C


ATOM
2728
NH1
ARG
A
196
−65.938
24.014
−2.937
1.00
19.84
N


ATOM
2731
NH2
ARG
A
196
−67.582
22.844
−3.999
1.00
18.53
N


ATOM
2734
C
ARG
A
196
−65.070
22.982
2.061
1.00
19.02
C


ATOM
2735
O
ARG
A
196
−66.276
23.085
2.284
1.00
19.30
O


ATOM
2737
N
LEU
A
197
−64.127
23.152
2.990
1.00
19.09
N


ATOM
2738
CA
LEU
A
197
−64.444
23.432
4.379
1.00
19.19
C


ATOM
2740
CB
LEU
A
197
−63.203
23.346
5.262
1.00
19.14
C


ATOM
2743
CG
LEU
A
197
−63.135
22.097
6.130
1.00
19.99
C


ATOM
2745
CD1
LEU
A
197
−64.476
21.899
6.811
1.00
21.56
C


ATOM
2749
CD2
LEU
A
197
−62.016
22.198
7.161
1.00
20.06
C


ATOM
2753
C
LEU
A
197
−65.071
24.781
4.553
1.00
19.52
C


ATOM
2754
O
LEU
A
197
−66.052
24.902
5.252
1.00
19.85
O


ATOM
2756
N
GLU
A
198
−64.488
25.803
3.941
1.00
20.11
N


ATOM
2757
CA
GLU
A
198
−65.051
27.147
3.986
1.00
20.47
C


ATOM
2759
CB
GLU
A
198
−64.079
28.156
3.385
1.00
20.84
C


ATOM
2762
CG
GLU
A
198
−64.659
29.539
3.069
1.00
22.30
C


ATOM
2765
CD
GLU
A
198
−65.206
30.260
4.283
1.00
24.40
C


ATOM
2766
OE1
GLU
A
198
−64.852
29.894
5.418
1.00
25.34
O


ATOM
2767
OE2
GLU
A
198
−65.991
31.217
4.106
1.00
26.83
O


ATOM
2768
C
GLU
A
198
−66.360
27.186
3.226
1.00
20.57
C


ATOM
2769
O
GLU
A
198
−67.212
28.009
3.529
1.00
20.65
O


ATOM
2771
N
ALA
A
199
−66.519
26.298
2.240
1.00
20.76
N


ATOM
2772
CA
ALA
A
199
−67.751
26.229
1.452
1.00
20.52
C


ATOM
2774
CB
ALA
A
199
−67.521
25.492
.166
1.00
20.44
C


ATOM
2778
C
ALA
A
199
−68.907
25.611
2.232
1.00
20.55
C


ATOM
2779
O
ALA
A
199
−69.988
26.158
2.202
1.00
20.17
O


ATOM
2781
N
VAL
A
200
−68.701
24.493
2.933
1.00
21.10
N


ATOM
2782
CA
VAL
A
200
−69.791
23.935
3.774
1.00
21.51
C


ATOM
2784
CB
VAL
A
200
−69.509
22.576
4.485
1.00
21.18
C


ATOM
2786
CG1
VAL
A
200
−69.618
21.460
3.525
1.00
21.27
C


ATOM
2790
CG2
VAL
A
200
−68.161
22.569
5.200
1.00
21.23
C


ATOM
2794
C
VAL
A
200
−70.170
24.905
4.860
1.00
21.99
C


ATOM
2795
O
VAL
A
200
−71.338
25.022
5.186
1.00
22.99
O


ATOM
2797
N
TRP
A
201
−69.204
25.590
5.440
1.00
22.10
N


ATOM
2798
CA
TRP
A
201
−69.551
26.565
6.433
1.00
22.58
C


ATOM
2800
CB
TRP
A
201
−68.315
27.144
7.121
1.00
22.92
C


ATOM
2803
CG
TRP
A
201
−68.667
27.957
8.310
1.00
23.65
C


ATOM
2804
CD1
TRP
A
201
−68.758
27.520
9.588
1.00
25.08
C


ATOM
2806
NE1
TRP
A
201
−69.127
28.549
10.417
1.00
26.35
N


ATOM
2808
CE2
TRP
A
201
−69.290
29.683
9.669
1.00
25.48
C


ATOM
2809
CD2
TRP
A
201
−69.010
29.344
8.332
1.00
24.77
C


ATOM
2810
CE3
TRP
A
201
−69.095
30.336
7.355
1.00
24.92
C


ATOM
2812
CZ3
TRP
A
201
−69.450
31.616
7.739
1.00
24.96
C


ATOM
2814
CH2
TRP
A
201
−69.720
31.918
9.078
1.00
25.08
C


ATOM
2816
CZ2
TRP
A
201
−69.648
30.968
10.056
1.00
24.68
C


ATOM
2818
C
TRP
A
201
−70.351
27.662
5.758
1.00
22.71
C


ATOM
2819
O
TRP
A
201
−71.482
27.921
6.140
1.00
22.76
O


ATOM
2821
N
SER
A
202
−69.776
28.272
4.727
1.00
23.04
N


ATOM
2822
CA
SER
A
202
−70.335
29.500
4.165
1.00
23.29
C


ATOM
2824
CB
SER
A
202
−69.304
30.205
3.290
1.00
23.20
C


ATOM
2827
OG
SER
A
202
−68.306
30.807
4.111
1.00
23.07
O


ATOM
2829
C
SER
A
202
−71.666
29.341
3.432
1.00
23.66
C


ATOM
2830
O
SER
A
202
−72.392
30.310
3.265
1.00
23.38
O


ATOM
2832
N
ILE
A
203
−71.996
28.124
3.016
1.00
24.49
N


ATOM
2833
CA
ILE
A
203
−73.321
27.859
2.460
1.00
25.07
C


ATOM
2835
CB
ILE
A
203
−73.391
26.530
1.653
1.00
24.74
C


ATOM
2837
CG1
ILE
A
203
−72.494
26.587
.423
1.00
24.36
C


ATOM
2840
CD1
ILE
A
203
−72.344
25.255
−.281
1.00
23.81
C


ATOM
2844
CG2
ILE
A
203
−74.795
26.292
1.156
1.00
24.45
C


ATOM
2848
C
ILE
A
203
−74.338
27.858
3.607
1.00
25.91
C


ATOM
2849
O
ILE
A
203
−75.350
28.554
3.546
1.00
26.26
O


ATOM
2851
N
GLU
A
204
−74.051
27.104
4.662
1.00
26.69
N


ATOM
2852
CA
GLU
A
204
−74.929
27.053
5.824
1.00
27.40
C


ATOM
2854
CB
GLU
A
204
−74.286
26.217
6.931
1.00
27.65
C


ATOM
2857
CG
GLU
A
204
−75.132
26.032
8.183
1.00
29.02
C


ATOM
2860
CD
GLU
A
204
−76.530
25.491
7.903
1.00
30.44
C


ATOM
2861
OE1
GLU
A
204
−76.746
24.858
6.844
1.00
31.22
O


ATOM
2862
OE2
GLU
A
204
−77.417
25.703
8.753
1.00
31.96
O


ATOM
2863
C
GLU
A
204
−75.258
28.451
6.342
1.00
27.61
C


ATOM
2864
O
GLU
A
204
−76.409
28.778
6.526
1.00
27.78
O


ATOM
2866
N
ALA
A
205
−74.240
29.274
6.547
1.00
28.15
N


ATOM
2867
CA
ALA
A
205
−74.416
30.637
7.050
1.00
28.41
C


ATOM
2869
CB
ALA
A
205
−73.062
31.285
7.274
1.00
28.41
C


ATOM
2873
C
ALA
A
205
−75.234
31.520
6.136
1.00
28.84
C


ATOM
2874
O
ALA
A
205
−76.050
32.302
6.610
1.00
28.54
O


ATOM
2876
N
TYR
A
206
−74.978
31.408
4.832
1.00
29.84
N


ATOM
2877
CA
TYR
A
206
−75.581
32.281
3.805
1.00
30.41
C


ATOM
2879
CB
TYR
A
206
−74.801
32.170
2.493
1.00
30.20
C


ATOM
2882
CG
TYR
A
206
−75.180
33.173
1.425
1.00
29.62
C


ATOM
2883
CD1
TYR
A
206
−74.721
34.482
1.484
1.00
30.32
C


ATOM
2885
CE1
TYR
A
206
−75.047
35.412
.489
1.00
29.70
C


ATOM
2887
CZ
TYR
A
206
−75.830
35.028
−.578
1.00
28.41
C


ATOM
2888
OH
TYR
A
206
−76.155
35.956
−1.548
1.00
26.74
O


ATOM
2890
CE2
TYR
A
206
−76.291
33.724
−.659
1.00
28.16
C


ATOM
2892
CD2
TYR
A
206
−75.958
32.806
.335
1.00
28.39
C


ATOM
2894
C
TYR
A
206
−77.038
31.918
3.561
1.00
31.15
C


ATOM
2895
O
TYR
A
206
−77.858
32.781
3.268
1.00
31.30
O


ATOM
2897
N
ARG
A
207
−77.341
30.631
3.676
1.00
32.05
N


ATOM
2898
CA
ARG
A
207
−78.691
30.128
3.555
1.00
32.81
C


ATOM
2900
CB
ARG
A
207
−78.684
28.623
3.813
1.00
32.86
C


ATOM
2903
CG
ARG
A
207
−80.028
27.941
3.673
1.00
34.01
C


ATOM
2906
CD
ARG
A
207
−80.066
26.632
4.450
1.00
35.11
C


ATOM
2909
NE
ARG
A
207
−79.096
25.669
3.939
1.00
35.35
N


ATOM
2911
CZ
ARG
A
207
−79.353
24.737
3.022
1.00
36.00
C


ATOM
2912
NH1
ARG
A
207
−80.562
24.611
2.478
1.00
35.55
N


ATOM
2915
NH2
ARG
A
207
−78.381
23.918
2.640
1.00
37.05
N


ATOM
2918
C
ARG
A
207
−79.609
30.826
4.550
1.00
33.57
C


ATOM
2919
O
ARG
A
207
−80.783
31.003
4.263
1.00
33.82
O


ATOM
2921
N
LYS
A
208
−79.064
31.223
5.707
1.00
34.58
N


ATOM
2922
CA
LYS
A
208
−79.835
31.830
6.812
1.00
35.21
C


ATOM
2924
CB
LYS
A
208
−79.137
31.599
8.159
1.00
35.05
C


ATOM
2927
CG
LYS
A
208
−78.847
30.147
8.496
1.00
34.78
C


ATOM
2930
CD
LYS
A
208
−78.583
29.974
9.980
1.00
34.44
C


ATOM
2933
CE
LYS
A
208
−77.992
28.619
10.309
1.00
34.16
C


ATOM
2936
NZ
LYS
A
208
−76.540
28.693
10.572
1.00
33.81
N


ATOM
2940
C
LYS
A
208
−80.075
33.333
6.663
1.00
36.06
C


ATOM
2941
O
LYS
A
208
−81.032
33.857
7.226
1.00
36.19
O


ATOM
2943
N
LYS
A
209
−79.197
34.032
5.946
1.00
36.93
N


ATOM
2944
CA
LYS
A
209
−79.412
35.445
5.652
1.00
37.77
C


ATOM
2946
CB
LYS
A
209
−78.284
35.998
4.779
1.00
38.18
C


ATOM
2949
CG
LYS
A
209
−76.928
36.134
5.458
1.00
39.90
C


ATOM
2952
CD
LYS
A
209
−75.982
37.023
4.621
1.00
42.39
C


ATOM
2955
CE
LYS
A
209
−74.531
37.016
5.169
1.00
44.21
C


ATOM
2958
NZ
LYS
A
209
−74.350
37.688
6.509
1.00
44.75
N


ATOM
2962
C
LYS
A
209
−80.738
35.600
4.911
1.00
37.89
C


ATOM
2963
O
LYS
A
209
−81.049
34.785
4.036
1.00
38.20
O


ATOM
2965
N
GLU
A
210
−81.518
36.632
5.243
1.00
37.87
N


ATOM
2966
CA
GLU
A
210
−82.847
36.797
4.630
1.00
37.72
C


ATOM
2968
CB
GLU
A
210
−83.821
37.578
5.549
1.00
38.19
C


ATOM
2971
CG
GLU
A
210
−83.850
39.130
5.404
1.00
39.68
C


ATOM
2974
CD
GLU
A
210
−85.270
39.736
5.534
1.00
41.27
C


ATOM
2975
OE1
GLU
A
210
−86.172
39.100
6.136
1.00
41.67
O


ATOM
2976
OE2
GLU
A
210
−85.482
40.859
5.019
1.00
42.17
O


ATOM
2977
C
GLU
A
210
−82.760
37.394
3.219
1.00
36.68
C


ATOM
2978
O
GLU
A
210
−83.605
37.103
2.371
1.00
36.39
O


ATOM
2980
N
ASP
A
211
−81.725
38.198
2.969
1.00
35.65
N


ATOM
2981
CA
ASP
A
211
−81.481
38.777
1.632
1.00
34.94
C


ATOM
2983
CB
ASP
A
211
−81.064
40.257
1.740
1.00
35.22
C


ATOM
2986
CG
ASP
A
211
−79.811
40.477
2.599
1.00
35.84
C


ATOM
2987
OD1
ASP
A
211
−79.379
39.557
3.341
1.00
35.60
O


ATOM
2988
OD2
ASP
A
211
−79.276
41.606
2.536
1.00
36.73
O


ATOM
2989
C
ASP
A
211
−80.451
37.989
.811
1.00
33.73
C


ATOM
2990
O
ASP
A
211
−79.753
38.556
−.021
1.00
33.40
O


ATOM
2992
N
ALA
A
212
−80.366
36.682
1.057
1.00
32.53
N


ATOM
2993
CA
ALA
A
212
−79.506
35.783
.293
1.00
31.40
C


ATOM
2995
CB
ALA
A
212
−79.569
34.382
.889
1.00
30.79
C


ATOM
2999
C
ALA
A
212
−79.940
35.762
−1.176
1.00
30.60
C


ATOM
3000
O
ALA
A
212
−81.124
35.594
−1.467
1.00
30.37
O


ATOM
3002
N
ASN
A
213
−79.000
35.967
−2.097
1.00
29.71
N


ATOM
3003
CA
ASN
A
213
−79.304
35.809
−3.520
1.00
29.27
C


ATOM
3005
CB
ASN
A
213
−78.144
36.315
−4.402
1.00
29.30
C


ATOM
3008
CG
ASN
A
213
−78.442
36.217
−5.911
1.00
29.07
C


ATOM
3009
OD1
ASN
A
213
−78.437
37.215
−6.624
1.00
29.38
O


ATOM
3010
ND2
ASN
A
213
−78.691
35.015
−6.389
1.00
28.37
N


ATOM
3013
C
ASN
A
213
−79.634
34.328
−3.785
1.00
28.81
C


ATOM
3014
O
ASN
A
213
−78.784
33.455
−3.658
1.00
29.04
O


ATOM
3016
N
GLN
A
214
−80.880
34.053
−4.145
1.00
28.04
N


ATOM
3017
CA
GLN
A
214
−81.376
32.692
−4.191
1.00
27.32
C


ATOM
3019
CB
GLN
A
214
−82.906
32.700
−4.178
1.00
27.47
C


ATOM
3022
CG
GLN
A
214
−83.505
33.269
−2.889
1.00
27.68
C


ATOM
3025
CD
GLN
A
214
−83.294
32.359
−1.688
1.00
27.54
C


ATOM
3026
OE1
GLN
A
214
−83.940
31.320
−1.571
1.00
28.19
O


ATOM
3027
NE2
GLN
A
214
−82.392
32.748
−.790
1.00
26.90
N


ATOM
3030
C
GLN
A
214
−80.858
31.902
−5.385
1.00
26.71
C


ATOM
3031
O
GLN
A
214
−80.863
30.668
−5.355
1.00
26.91
O


ATOM
3033
N
VAL
A
215
−80.439
32.600
−6.442
1.00
25.81
N


ATOM
3034
CA
VAL
A
215
−79.789
31.947
−7.588
1.00
24.88
C


ATOM
3036
CB
VAL
A
215
−79.651
32.894
−8.808
1.00
24.81
C


ATOM
3038
CG1
VAL
A
215
−78.459
32.483
−9.680
1.00
24.94
C


ATOM
3042
CG2
VAL
A
215
−80.935
32.907
−9.620
1.00
23.74
C


ATOM
3046
C
VAL
A
215
−78.415
31.427
−7.176
1.00
24.18
C


ATOM
3047
O
VAL
A
215
−78.073
30.282
−7.430
1.00
23.74
O


ATOM
3049
N
LEU
A
216
−77.650
32.283
−6.510
1.00
23.78
N


ATOM
3050
CA
LEU
A
216
−76.299
31.961
−6.036
1.00
23.33
C


ATOM
3052
CB
LEU
A
216
−75.680
33.195
−5.373
1.00
23.10
C


ATOM
3055
CG
LEU
A
216
−74.194
33.192
−5.059
1.00
22.91
C


ATOM
3057
CD1
LEU
A
216
−73.383
32.962
−6.312
1.00
22.88
C


ATOM
3061
CD2
LEU
A
216
−73.826
34.518
−4.424
1.00
22.03
C


ATOM
3065
C
LEU
A
216
−76.333
30.797
−5.050
1.00
22.92
C


ATOM
3066
O
LEU
A
216
−75.598
29.826
−5.201
1.00
23.38
O


ATOM
3068
N
LEU
A
217
−77.214
30.890
−4.060
1.00
22.09
N


ATOM
3069
CA
LEU
A
217
−77.367
29.851
−3.054
1.00
21.14
C


ATOM
3071
CB
LEU
A
217
−78.488
30.225
−2.090
1.00
21.11
C


ATOM
3074
CG
LEU
A
217
−78.782
29.236
−.961
1.00
21.08
C


ATOM
3076
CD1
LEU
A
217
−77.703
29.267
.101
1.00
19.87
C


ATOM
3080
CD2
LEU
A
217
−80.146
29.535
−.360
1.00
21.46
C


ATOM
3084
C
LEU
A
217
−77.682
28.517
−3.685
1.00
20.38
C


ATOM
3085
O
LEU
A
217
−77.133
27.508
−3.286
1.00
20.04
O


ATOM
3087
N
GLU
A
218
−78.579
28.512
−4.661
1.00
19.97
N


ATOM
3088
CA
GLU
A
218
−79.027
27.260
−5.273
1.00
19.72
C


ATOM
3090
CB
GLU
A
218
−80.220
27.499
−6.212
1.00
19.87
C


ATOM
3093
CG
GLU
A
218
−81.005
26.228
−6.627
1.00
21.03
C


ATOM
3096
CD
GLU
A
218
−82.303
26.535
−7.424
1.00
22.59
C


ATOM
3097
OE1
GLU
A
218
−82.643
27.730
−7.619
1.00
23.61
O


ATOM
3098
OE2
GLU
A
218
−82.983
25.578
−7.863
1.00
22.31
O


ATOM
3099
C
GLU
A
218
−77.854
26.613
−6.009
1.00
19.03
C


ATOM
3100
O
GLU
A
218
−77.522
25.452
−5.764
1.00
18.97
O


ATOM
3102
N
LEU
A
219
−77.203
27.381
−6.878
1.00
18.06
N


ATOM
3103
CA
LEU
A
219
−76.012
26.907
−7.573
1.00
17.15
C


ATOM
3105
CB
LEU
A
219
−75.439
28.029
−8.418
1.00
16.75
C


ATOM
3108
CG
LEU
A
219
−74.196
27.717
−9.231
1.00
16.01
C


ATOM
3110
CD1
LEU
A
219
−74.511
26.832
−10.404
1.00
12.99
C


ATOM
3114
CD2
LEU
A
219
−73.577
29.028
−9.675
1.00
16.20
C


ATOM
3118
C
LEU
A
219
−74.973
26.409
−6.570
1.00
16.75
C


ATOM
3119
O
LEU
A
219
−74.377
25.355
−6.757
1.00
16.47
O


ATOM
3121
N
ALA
A
220
−74.786
27.169
−5.496
1.00
16.40
N


ATOM
3122
CA
ALA
A
220
−73.858
26.808
−4.422
1.00
16.15
C


ATOM
3124
CB
ALA
A
220
−73.906
27.830
−3.302
1.00
16.13
C


ATOM
3128
C
ALA
A
220
−74.137
25.436
−3.862
1.00
16.03
C


ATOM
3129
O
ALA
A
220
−73.253
24.603
−3.799
1.00
16.30
O


ATOM
3131
N
ILE
A
221
−75.371
25.201
−3.447
1.00
16.24
N


ATOM
3132
CA
ILE
A
221
−75.731
23.916
−2.857
1.00
16.33
C


ATOM
3134
CB
ILE
A
221
−77.203
23.895
−2.352
1.00
15.94
C


ATOM
3136
CG1
ILE
A
221
−77.410
24.879
−1.203
1.00
15.36
C


ATOM
3139
CD1
ILE
A
221
−78.842
25.234
−.962
1.00
14.49
C


ATOM
3143
CG2
ILE
A
221
−77.571
22.537
−1.836
1.00
15.18
C


ATOM
3147
C
ILE
A
221
−75.509
22.841
−3.911
1.00
17.06
C


ATOM
3148
O
ILE
A
221
−74.899
21.819
−3.653
1.00
16.51
O


ATOM
3150
N
LEU
A
222
−75.981
23.138
−5.117
1.00
18.53
N


ATOM
3151
CA
LEU
A
222
−76.006
22.207
−6.243
1.00
19.25
C


ATOM
3153
CB
LEU
A
222
−76.657
22.891
−7.445
1.00
19.18
C


ATOM
3156
CG
LEU
A
222
−77.037
21.972
−8.597
1.00
19.65
C


ATOM
3158
CD1
LEU
A
222
−78.318
22.489
−9.268
1.00
19.24
C


ATOM
3162
CD2
LEU
A
222
−75.869
21.788
−9.597
1.00
18.75
C


ATOM
3166
C
LEU
A
222
−74.618
21.751
−6.630
1.00
20.07
C


ATOM
3167
O
LEU
A
222
−74.381
20.558
−6.778
1.00
20.55
O


ATOM
3169
N
ASP
A
223
−73.710
22.711
−6.797
1.00
20.92
N


ATOM
3170
CA
ASP
A
223
−72.330
22.427
−7.189
1.00
21.43
C


ATOM
3172
CB
ASP
A
223
−71.621
23.723
−7.585
1.00
21.63
C


ATOM
3175
CG
ASP
A
223
−70.202
23.495
−8.070
1.00
22.51
C


ATOM
3176
OD1
ASP
A
223
−69.296
23.282
−7.225
1.00
24.67
O


ATOM
3177
OD2
ASP
A
223
−69.990
23.544
−9.296
1.00
22.73
O


ATOM
3178
C
ASP
A
223
−71.573
21.722
−6.060
1.00
21.79
C


ATOM
3179
O
ASP
A
223
−70.843
20.764
−6.313
1.00
21.73
O


ATOM
3181
N
TYR
A
224
−71.755
22.182
−4.820
1.00
22.14
N


ATOM
3182
CA
TYR
A
224
−71.120
21.529
−3.686
1.00
22.22
C


ATOM
3184
CB
TYR
A
224
−71.462
22.190
−2.355
1.00
22.38
C


ATOM
3187
CG
TYR
A
224
−70.673
21.542
−1.249
1.00
22.65
C


ATOM
3188
CD1
TYR
A
224
−69.356
21.864
−1.053
1.00
22.59
C


ATOM
3190
CE1
TYR
A
224
−68.622
21.258
−.098
1.00
23.38
C


ATOM
3192
CZ
TYR
A
224
−69.177
20.288
.670
1.00
23.22
C


ATOM
3193
OH
TYR
A
224
−68.402
19.684
1.620
1.00
23.93
O


ATOM
3195
CE2
TYR
A
224
−70.483
19.931
.500
1.00
23.36
C


ATOM
3197
CD2
TYR
A
224
−71.221
20.547
−.464
1.00
23.41
C


ATOM
3199
C
TYR
A
224
−71.485
20.065
−3.605
1.00
22.52
C


ATOM
3200
O
TYR
A
224
−70.641
19.233
−3.311
1.00
22.93
O


ATOM
3202
N
ASN
A
225
−72.742
19.749
−3.856
1.00
22.94
N


ATOM
3203
CA
ASN
A
225
−73.187
18.374
−3.801
1.00
23.44
C


ATOM
3205
CB
ASN
A
225
−74.706
18.310
−3.729
1.00
23.34
C


ATOM
3208
CG
ASN
A
225
−75.210
18.579
−2.352
1.00
22.62
C


ATOM
3209
OD1
ASN
A
225
−74.749
17.977
−1.407
1.00
23.33
O


ATOM
3210
ND2
ASN
A
225
−76.146
19.494
−2.223
1.00
22.52
N


ATOM
3213
C
ASN
A
225
−72.694
17.523
−4.959
1.00
24.26
C


ATOM
3214
O
ASN
A
225
−72.408
16.328
−4.767
1.00
24.88
O


ATOM
3216
N
MET
A
226
−72.607
18.119
−6.150
1.00
24.67
N


ATOM
3217
CA
MET
A
226
−72.167
17.395
−7.349
1.00
24.95
C


ATOM
3219
CB
MET
A
226
−72.421
18.237
−8.594
1.00
25.37
C


ATOM
3222
CG
MET
A
226
−71.785
17.713
−9.873
1.00
27.23
C


ATOM
3225
SD
MET
A
226
−70.837
19.010
−10.708
1.00
31.73
S


ATOM
3226
CE
MET
A
226
−72.185
20.000
−11.367
1.00
31.30
C


ATOM
3230
C
MET
A
226
−70.688
17.045
−7.247
1.00
24.70
C


ATOM
3231
O
MET
A
226
−70.291
15.929
−7.568
1.00
24.91
O


ATOM
3233
N
ILE
A
227
−69.873
17.995
−6.799
1.00
24.43
N


ATOM
3234
CA
ILE
A
227
−68.456
17.726
−6.598
1.00
24.18
C


ATOM
3236
CB
ILE
A
227
−67.656
18.971
−6.154
1.00
24.15
C


ATOM
3238
CG1
ILE
A
227
−67.628
20.021
−7.253
1.00
23.64
C


ATOM
3241
CD1
ILE
A
227
−66.853
21.246
−6.880
1.00
23.11
C


ATOM
3245
CG2
ILE
A
227
−66.229
18.603
−5.827
1.00
24.36
C


ATOM
3249
C
ILE
A
227
−68.329
16.645
−5.546
1.00
23.98
C


ATOM
3250
O
ILE
A
227
−67.609
15.689
−5.747
1.00
24.35
O


ATOM
3252
N
GLN
A
228
−69.048
16.779
−4.439
1.00
23.79
N


ATOM
3253
CA
GLN
A
228
−69.020
15.754
−3.382
1.00
23.66
C


ATOM
3255
CB
GLN
A
228
−70.077
16.010
−2.305
1.00
23.64
C


ATOM
3258
CG
GLN
A
228
−69.972
15.049
−1.145
1.00
22.91
C


ATOM
3261
CD
GLN
A
228
−70.891
15.417
−.033
1.00
23.26
C


ATOM
3262
OE1
GLN
A
228
−72.082
15.098
−.069
1.00
25.64
O


ATOM
3263
NE2
GLN
A
228
−70.357
16.083
.978
1.00
21.77
N


ATOM
3266
C
GLN
A
228
−69.243
14.349
−3.893
1.00
23.54
C


ATOM
3267
O
GLN
A
228
−68.664
13.412
−3.357
1.00
23.59
O


ATOM
3269
N
SER
A
229
−70.105
14.194
−4.893
1.00
23.27
N


ATOM
3270
CA
SER
A
229
−70.394
12.868
−5.406
1.00
23.35
C


ATOM
3272
CB
SER
A
229
−71.753
12.812
−6.095
1.00
23.31
C


ATOM
3275
OG
SER
A
229
−71.836
13.823
−7.060
1.00
24.31
O


ATOM
3277
C
SER
A
229
−69.287
12.394
−6.331
1.00
23.19
C


ATOM
3278
O
SER
A
229
−69.130
11.194
−6.512
1.00
23.44
O


ATOM
3280
N
VAL
A
230
−68.512
13.306
−6.914
1.00
23.14
N


ATOM
3281
CA
VAL
A
230
−67.269
12.874
−7.552
1.00
23.00
C


ATOM
3283
CB
VAL
A
230
−66.469
13.998
−8.253
1.00
22.75
C


ATOM
3285
CG1
VAL
A
230
−65.091
13.470
−8.667
1.00
21.87
C


ATOM
3289
CG2
VAL
A
230
−67.222
14.539
−9.459
1.00
21.74
C


ATOM
3293
C
VAL
A
230
−66.417
12.268
−6.452
1.00
23.35
C


ATOM
3294
O
VAL
A
230
−65.917
11.173
−6.589
1.00
23.68
O


ATOM
3296
N
TYR
A
231
−66.284
12.968
−5.340
1.00
23.96
N


ATOM
3297
CA
TYR
A
231
−65.414
12.509
−4.274
1.00
24.60
C


ATOM
3299
CB
TYR
A
231
−65.486
13.426
−3.051
1.00
24.69
C


ATOM
3302
CG
TYR
A
231
−64.963
14.837
−3.211
1.00
24.11
C


ATOM
3303
CD1
TYR
A
231
−64.071
15.190
−4.214
1.00
24.31
C


ATOM
3305
CE1
TYR
A
231
−63.592
16.491
−4.322
1.00
24.63
C


ATOM
3307
CZ
TYR
A
231
−63.994
17.441
−3.406
1.00
25.13
C


ATOM
3308
OH
TYR
A
231
−63.554
18.754
−3.456
1.00
24.64
O


ATOM
3310
CE2
TYR
A
231
−64.863
17.086
−2.402
1.00
25.76
C


ATOM
3312
CD2
TYR
A
231
−65.329
15.800
−2.308
1.00
24.28
C


ATOM
3314
C
TYR
A
231
−65.785
11.111
−3.833
1.00
25.26
C


ATOM
3315
O
TYR
A
231
−64.917
10.315
−3.488
1.00
25.44
O


ATOM
3317
N
GLN
A
232
−67.079
10.819
−3.833
1.00
26.04
N


ATOM
3318
CA
GLN
A
232
−67.566
9.532
−3.362
1.00
26.57
C


ATOM
3320
CB
GLN
A
232
−69.060
9.621
−3.057
1.00
26.44
C


ATOM
3323
CG
GLN
A
232
−69.339
10.397
−1.778
1.00
26.25
C


ATOM
3326
CD
GLN
A
232
−70.786
10.819
−1.630
1.00
26.53
C


ATOM
3327
OE1
GLN
A
232
−71.667
10.305
−2.318
1.00
27.77
O


ATOM
3328
NE2
GLN
A
232
−71.040
11.759
−.723
1.00
25.51
N


ATOM
3331
C
GLN
A
232
−67.238
8.414
−4.351
1.00
27.44
C


ATOM
3332
O
GLN
A
232
−66.852
7.319
−3.935
1.00
27.33
O


ATOM
3334
N
ARG
A
233
−67.373
8.691
−5.649
1.00
28.66
N


ATOM
3335
CA
ARG
A
233
−66.905
7.768
−6.685
1.00
29.86
C


ATOM
3337
CB
ARG
A
233
−67.212
8.292
−8.090
1.00
30.08
C


ATOM
3340
CG
ARG
A
233
−66.378
7.634
−9.179
1.00
32.80
C


ATOM
3343
CD
ARG
A
233
−66.913
7.923
−10.572
1.00
36.74
C


ATOM
3346
NE
ARG
A
233
−66.962
9.363
−10.860
1.00
40.33
N


ATOM
3348
CZ
ARG
A
233
−68.069
10.119
−10.891
1.00
43.12
C


ATOM
3349
NH1
ARG
A
233
−69.276
9.600
−10.655
1.00
44.55
N


ATOM
3352
NH2
ARG
A
233
−67.971
11.418
−11.169
1.00
43.38
N


ATOM
3355
C
ARG
A
233
−65.401
7.518
−6.514
1.00
30.34
C


ATOM
3356
O
ARG
A
233
−64.962
6.374
−6.412
1.00
30.55
O


ATOM
3358
N
ASP
A
234
−64.621
8.591
−6.461
1.00
31.00
N


ATOM
3359
CA
ASP
A
234
−63.187
8.487
−6.205
1.00
31.48
C


ATOM
3361
CB
ASP
A
234
−62.594
9.870
−5.942
1.00
31.56
C


ATOM
3364
CG
ASP
A
234
−62.573
10.741
−7.167
1.00
32.78
C


ATOM
3365
OD1
ASP
A
234
−62.911
10.249
−8.276
1.00
34.25
O


ATOM
3366
OD2
ASP
A
234
−62.215
11.930
−7.011
1.00
34.81
O


ATOM
3367
C
ASP
A
234
−62.885
7.596
−5.010
1.00
31.70
C


ATOM
3368
O
ASP
A
234
−61.985
6.780
−5.053
1.00
31.56
O


ATOM
3370
N
LEU
A
235
−63.647
7.766
−3.943
1.00
32.43
N


ATOM
3371
CA
LEU
A
235
−63.366
7.102
−2.685
1.00
33.04
C


ATOM
3373
CB
LEU
A
235
−64.072
7.833
−1.553
1.00
32.57
C


ATOM
3376
CG
LEU
A
235
−63.884
7.252
−.167
1.00
30.85
C


ATOM
3378
CD1
LEU
A
235
−62.428
7.261
.174
1.00
28.78
C


ATOM
3382
CD2
LEU
A
235
−64.703
8.050
.823
1.00
29.93
C


ATOM
3386
C
LEU
A
235
−63.789
5.637
−2.686
1.00
34.69
C


ATOM
3387
O
LEU
A
235
−63.123
4.812
−2.065
1.00
35.29
O


ATOM
3389
N
ARG
A
236
−64.893
5.299
−3.354
1.00
36.09
N


ATOM
3390
CA
ARG
A
236
−65.290
3.893
−3.458
1.00
37.26
C


ATOM
3392
CB
ARG
A
236
−66.672
3.736
−4.103
1.00
37.64
C


ATOM
3395
CG
ARG
A
236
−67.839
4.000
−3.157
1.00
39.15
C


ATOM
3398
CD
ARG
A
236
−69.162
3.528
−3.741
1.00
40.25
C


ATOM
3401
NE
ARG
A
236
−69.385
4.046
−5.095
1.00
41.64
N


ATOM
3403
CZ
ARG
A
236
−69.837
5.269
−5.392
1.00
42.54
C


ATOM
3404
NH1
ARG
A
236
−70.115
6.147
−4.430
1.00
42.18
N


ATOM
3407
NH2
ARG
A
236
−70.012
5.621
−6.668
1.00
43.04
N


ATOM
3410
C
ARG
A
236
−64.266
3.108
−4.262
1.00
37.81
C


ATOM
3411
O
ARG
A
236
−64.104
1.912
−4.060
1.00
38.07
O


ATOM
3413
N
GLU
A
237
−63.584
3.801
−5.168
1.00
38.69
N


ATOM
3414
CA
GLU
A
237
−62.639
3.189
−6.102
1.00
39.45
C


ATOM
3416
CB
GLU
A
237
−62.452
4.108
−7.338
1.00
40.25
C


ATOM
3419
CG
GLU
A
237
−62.268
3.387
−8.710
1.00
43.02
C


ATOM
3422
CD
GLU
A
237
−60.835
3.458
−9.287
1.00
46.64
C


ATOM
3423
OE1
GLU
A
237
−60.093
4.440
−8.997
1.00
48.03
O


ATOM
3424
OE2
GLU
A
237
−60.466
2.519
−10.047
1.00
48.35
O


ATOM
3425
C
GLU
A
237
−61.310
2.905
−5.408
1.00
38.70
C


ATOM
3426
O
GLU
A
237
−60.832
1.782
−5.440
1.00
38.33
O


ATOM
3428
N
THR
A
238
−60.726
3.908
−4.764
1.00
38.40
N


ATOM
3429
CA
THR
A
238
−59.504
3.669
−4.009
1.00
38.74
C


ATOM
3431
CB
THR
A
238
−58.710
4.965
−3.593
1.00
38.78
C


ATOM
3433
OG1
THR
A
238
−59.294
5.583
−2.444
1.00
38.38
O


ATOM
3435
CG2
THR
A
238
−58.612
5.963
−4.743
1.00
38.94
C


ATOM
3439
C
THR
A
238
−59.802
2.834
−2.773
1.00
39.05
C


ATOM
3440
O
THR
A
238
−58.886
2.294
−2.153
1.00
39.17
O


ATOM
3442
N
SER
A
239
−61.076
2.717
−2.408
1.00
39.34
N


ATOM
3443
CA
SER
A
239
−61.443
1.841
−1.304
1.00
39.47
C


ATOM
3445
CB
SER
A
239
−62.844
2.149
−.792
1.00
39.34
C


ATOM
3448
OG
SER
A
239
−63.087
1.428
.396
1.00
39.94
O


ATOM
3450
C
SER
A
239
−61.313
.367
−1.699
1.00
39.53
C


ATOM
3451
O
SER
A
239
−60.834
−.439
−.900
1.00
39.53
O


ATOM
3453
N
ARG
A
240
−61.728
.021
−2.921
1.00
39.69
N


ATOM
3454
CA
ARG
A
240
−61.559
−1.341
−3.439
1.00
39.79
C


ATOM
3456
CB
ARG
A
240
−62.105
−1.492
−4.867
1.00
40.34
C


ATOM
3459
CG
ARG
A
240
−63.624
−1.715
−4.947
1.00
43.03
C


ATOM
3462
CD
ARG
A
240
−64.104
−2.125
−6.364
1.00
46.66
C


ATOM
3465
NE
ARG
A
240
−63.780
−1.136
−7.416
1.00
50.46
N


ATOM
3467
CZ
ARG
A
240
−64.535
−.082
−7.775
1.00
53.13
C


ATOM
3468
NH1
ARG
A
240
−64.112
.732
−8.749
1.00
53.13
N


ATOM
3471
NH2
ARG
A
240
−65.707
.178
−7.178
1.00
54.02
N


ATOM
3474
C
ARG
A
240
−60.089
−1.704
−3.414
1.00
38.94
C


ATOM
3475
O
ARG
A
240
−59.732
−2.795
−2.978
1.00
39.09
O


ATOM
3477
N
TRP
A
241
−59.246
−.775
−3.862
1.00
37.92
N


ATOM
3478
CA
TRP
A
241
−57.787
−.947
−3.842
1.00
36.93
C


ATOM
3480
CB
TRP
A
241
−57.099
.318
−4.388
1.00
36.57
C


ATOM
3483
CG
TRP
A
241
−55.624
.333
−4.209
1.00
34.71
C


ATOM
3484
CD1
TRP
A
241
−54.702
−.312
−4.969
1.00
33.50
C


ATOM
3486
NE1
TRP
A
241
−53.441
−.068
−4.482
1.00
32.62
N


ATOM
3488
CE2
TRP
A
241
−53.538
.747
−3.392
1.00
31.16
C


ATOM
3489
CD2
TRP
A
241
−54.896
1.022
−3.191
1.00
32.61
C


ATOM
3490
CE3
TRP
A
241
−55.268
1.837
−2.117
1.00
32.63
C


ATOM
3492
CZ3
TRP
A
241
−54.293
2.345
−1.307
1.00
31.31
C


ATOM
3494
CH2
TRP
A
241
−52.956
2.053
−1.537
1.00
31.81
C


ATOM
3496
CZ2
TRP
A
241
−52.560
1.252
−2.577
1.00
31.18
C


ATOM
3498
C
TRP
A
241
−57.263
−1.274
−2.444
1.00
36.67
C


ATOM
3499
O
TRP
A
241
−56.540
−2.247
−2.252
1.00
36.32
O


ATOM
3501
N
TRP
A
242
−57.647
−.460
−1.471
1.00
36.59
N


ATOM
3502
CA
TRP
A
242
−57.150
−.593
−.100
1.00
36.80
C


ATOM
3504
CB
TRP
A
242
−57.694
.582
.734
1.00
36.47
C


ATOM
3507
CG
TRP
A
242
−57.113
.754
2.107
1.00
35.59
C


ATOM
3508
CD1
TRP
A
242
−57.806
.831
3.270
1.00
35.14
C


ATOM
3510
NE1
TRP
A
242
−56.950
.999
4.328
1.00
34.47
N


ATOM
3512
CE2
TRP
A
242
−55.667
1.032
3.859
1.00
34.64
C


ATOM
3513
CD2
TRP
A
242
−55.728
.884
2.461
1.00
35.26
C


ATOM
3514
CE3
TRP
A
242
−54.531
.882
1.732
1.00
35.22
C


ATOM
3516
CZ3
TRP
A
242
−53.342
1.024
2.409
1.00
34.30
C


ATOM
3518
CH2
TRP
A
242
−53.318
1.169
3.799
1.00
34.70
C


ATOM
3520
CZ2
TRP
A
242
−54.467
1.173
4.542
1.00
34.63
C


ATOM
3522
C
TRP
A
242
−57.482
−1.975
.524
1.00
37.34
C


ATOM
3523
O
TRP
A
242
−56.628
−2.623
1.126
1.00
36.53
O


ATOM
3525
N
ARG
A
243
−58.720
−2.421
.348
1.00
38.54
N


ATOM
3526
CA
ARG
A
243
−59.149
−3.734
.822
1.00
39.71
C


ATOM
3528
CB
ARG
A
243
−60.669
−3.896
.687
1.00
40.10
C


ATOM
3531
CG
ARG
A
243
−61.495
−3.134
1.747
1.00
42.18
C


ATOM
3534
CD
ARG
A
243
−62.826
−2.623
1.169
1.00
45.18
C


ATOM
3537
NE
ARG
A
243
−63.506
−3.656
.369
1.00
48.22
N


ATOM
3539
CZ
ARG
A
243
−64.374
−3.431
−.629
1.00
49.95
C


ATOM
3540
NH1
ARG
A
243
−64.717
−2.182
−.991
1.00
50.13
N


ATOM
3543
NH2
ARG
A
243
−64.911
−4.477
−1.273
1.00
49.77
N


ATOM
3546
C
ARG
A
243
−58.438
−4.852
.068
1.00
40.08
C


ATOM
3547
O
ARG
A
243
−58.084
−5.870
.665
1.00
40.45
O


ATOM
3549
N
ARG
A
244
−58.236
−4.663
−1.236
1.00
40.42
N


ATOM
3550
CA
ARG
A
244
−57.499
−5.621
−2.063
1.00
40.65
C


ATOM
3552
CB
ARG
A
244
−57.370
−5.100
−3.503
1.00
41.29
C


ATOM
3555
CG
ARG
A
244
−56.939
−6.126
−4.560
1.00
43.27
C


ATOM
3558
CD
ARG
A
244
−58.090
−7.062
−4.952
1.00
46.01
C


ATOM
3561
NE
ARG
A
244
−57.595
−8.337
−5.485
1.00
48.77
N


ATOM
3563
CZ
ARG
A
244
−57.075
−9.332
−4.752
1.00
51.17
C


ATOM
3564
NH1
ARG
A
244
−56.968
−9.234
−3.422
1.00
51.63
N


ATOM
3567
NH2
ARG
A
244
−56.656
−10.448
−5.353
1.00
52.11
N


ATOM
3570
C
ARG
A
244
−56.120
−5.861
−1.465
1.00
40.12
C


ATOM
3571
O
ARG
A
244
−55.728
−7.000
−1.235
1.00
39.96
O


ATOM
3573
N
VAL
A
245
−55.399
−4.778
−1.201
1.00
39.78
N


ATOM
3574
CA
VAL
A
245
−54.099
−4.858
−.543
1.00
39.68
C


ATOM
3576
CB
VAL
A
245
−53.430
−3.458
−.437
1.00
39.70
C


ATOM
3578
CG1
VAL
A
245
−53.133
−2.914
−1.824
1.00
39.54
C


ATOM
3582
CG2
VAL
A
245
−52.145
−3.507
.395
1.00
39.41
C


ATOM
3586
C
VAL
A
245
−54.262
−5.472
.841
1.00
39.73
C


ATOM
3587
O
VAL
A
245
−53.455
−6.291
1.253
1.00
39.53
O


ATOM
3589
N
GLY
A
246
−55.312
−5.051
1.544
1.00
40.11
N


ATOM
3590
CA
GLY
A
246
−55.705
−5.617
2.839
1.00
40.35
C


ATOM
3593
C
GLY
A
246
−54.621
−5.648
3.894
1.00
40.58
C


ATOM
3594
O
GLY
A
246
−54.396
−6.676
4.510
1.00
40.68
O


ATOM
3596
N
LEU
A
247
−53.963
−4.522
4.129
1.00
41.15
N


ATOM
3597
CA
LEU
A
247
−52.778
−4.519
4.982
1.00
41.71
C


ATOM
3599
CB
LEU
A
247
−51.808
−3.422
4.537
1.00
41.60
C


ATOM
3602
CG
LEU
A
247
−50.334
−3.815
4.441
1.00
40.83
C


ATOM
3604
CD1
LEU
A
247
−50.158
−5.014
3.539
1.00
40.54
C


ATOM
3608
CD2
LEU
A
247
−49.534
−2.644
3.918
1.00
39.92
C


ATOM
3612
C
LEU
A
247
−53.136
−4.385
6.471
1.00
42.60
C


ATOM
3613
O
LEU
A
247
−52.588
−5.111
7.313
1.00
42.06
O


ATOM
3615
N
ALA
A
248
−54.064
−3.475
6.788
1.00
43.75
N


ATOM
3616
CA
ALA
A
248
−54.603
−3.352
8.162
1.00
44.52
C


ATOM
3618
CB
ALA
A
248
−55.712
−2.296
8.210
1.00
44.31
C


ATOM
3622
C
ALA
A
248
−55.129
−4.706
8.695
1.00
45.10
C


ATOM
3623
O
ALA
A
248
−54.969
−5.041
9.875
1.00
44.96
O


ATOM
3625
N
THR
A
249
−55.742
−5.477
7.802
1.00
45.91
N


ATOM
3626
CA
THR
A
249
−56.328
−6.768
8.154
1.00
46.50
C


ATOM
3628
CB
THR
A
249
−57.303
−7.277
7.041
1.00
46.48
C


ATOM
3630
OG1
THR
A
249
−56.583
−8.039
6.063
1.00
46.20
O


ATOM
3632
CG2
THR
A
249
−58.033
−6.102
6.350
1.00
46.70
C


ATOM
3636
C
THR
A
249
−55.257
−7.840
8.447
1.00
47.06
C


ATOM
3637
O
THR
A
249
−55.531
−8.813
9.159
1.00
47.22
O


ATOM
3639
N
LYS
A
250
−54.049
−7.661
7.905
1.00
47.52
N


ATOM
3640
CA
LYS
A
250
−52.952
−8.620
8.096
1.00
47.91
C


ATOM
3642
CB
LYS
A
250
−52.319
−8.976
6.744
1.00
48.09
C


ATOM
3645
CG
LYS
A
250
−52.911
−10.229
6.088
1.00
48.91
C


ATOM
3648
CD
LYS
A
250
−52.210
−11.510
6.585
1.00
49.88
C


ATOM
3651
CE
LYS
A
250
−53.066
−12.760
6.349
1.00
50.12
C


ATOM
3654
NZ
LYS
A
250
−52.385
−14.011
6.791
1.00
49.99
N


ATOM
3658
C
LYS
A
250
−51.880
−8.129
9.078
1.00
48.13
C


ATOM
3659
O
LYS
A
250
−51.252
−8.939
9.759
1.00
47.89
O


ATOM
3661
N
LEU
A
251
−51.667
−6.812
9.134
1.00
48.63
N


ATOM
3662
CA
LEU
A
251
−50.758
−6.187
10.114
1.00
48.83
C


ATOM
3664
CB
LEU
A
251
−49.981
−5.020
9.485
1.00
48.74
C


ATOM
3667
CG
LEU
A
251
−48.569
−5.296
8.960
1.00
48.51
C


ATOM
3669
CD1
LEU
A
251
−48.472
−6.594
8.174
1.00
47.96
C


ATOM
3673
CD2
LEU
A
251
−48.102
−4.115
8.116
1.00
48.76
C


ATOM
3677
C
LEU
A
251
−51.559
−5.703
11.324
1.00
49.17
C


ATOM
3678
O
LEU
A
251
−52.176
−4.632
11.304
1.00
49.28
O


ATOM
3680
N
HIS
A
252
−51.521
−6.486
12.393
1.00
49.53
N


ATOM
3681
CA
HIS
A
252
−52.470
−6.322
13.494
1.00
50.02
C


ATOM
3683
CB
HIS
A
252
−52.598
−7.647
14.266
1.00
50.42
C


ATOM
3686
CG
HIS
A
252
−52.860
−8.834
13.380
1.00
52.23
C


ATOM
3687
ND1
HIS
A
252
−54.032
−8.989
12.667
1.00
53.75
N


ATOM
3689
CE1
HIS
A
252
−53.978
−10.113
11.973
1.00
54.43
C


ATOM
3691
NE2
HIS
A
252
−52.810
−10.689
12.201
1.00
54.52
N


ATOM
3693
CD2
HIS
A
252
−52.090
−9.910
13.077
1.00
53.70
C


ATOM
3695
C
HIS
A
252
−52.161
−5.147
14.441
1.00
49.64
C


ATOM
3696
O
HIS
A
252
−52.951
−4.849
15.336
1.00
49.61
O


ATOM
3698
N
PHE
A
253
−51.027
−4.483
14.229
1.00
49.37
N


ATOM
3699
CA
PHE
A
253
−50.652
−3.273
14.973
1.00
49.15
C


ATOM
3701
CB
PHE
A
253
−49.144
−3.273
15.260
1.00
49.07
C


ATOM
3704
CG
PHE
A
253
−48.307
−3.112
14.025
1.00
48.30
C


ATOM
3705
CD1
PHE
A
253
−48.028
−1.851
13.518
1.00
48.10
C


ATOM
3707
CE1
PHE
A
253
−47.290
−1.702
12.349
1.00
48.21
C


ATOM
3709
CZ
PHE
A
253
−46.825
−2.825
11.675
1.00
47.88
C


ATOM
3711
CE2
PHE
A
253
−47.101
−4.090
12.175
1.00
47.66
C


ATOM
3713
CD2
PHE
A
253
−47.842
−4.226
13.338
1.00
47.74
C


ATOM
3715
C
PHE
A
253
−50.974
−2.006
14.176
1.00
49.31
C


ATOM
3716
O
PHE
A
253
−50.846
−.893
14.702
1.00
49.01
O


ATOM
3718
N
ALA
A
254
−51.365
−2.185
12.910
1.00
49.48
N


ATOM
3719
CA
ALA
A
254
−51.412
−1.095
11.931
1.00
49.63
C


ATOM
3721
CB
ALA
A
254
−51.344
−1.663
10.524
1.00
49.64
C


ATOM
3725
C
ALA
A
254
−52.650
−.224
12.061
1.00
49.77
C


ATOM
3726
O
ALA
A
254
−53.761
−.735
12.219
1.00
49.86
O


ATOM
3728
N
ARG
A
255
−52.452
1.090
11.975
1.00
49.93
N


ATOM
3729
CA
ARG
A
255
−53.562
2.042
11.908
1.00
50.27
C


ATOM
3731
CB
ARG
A
255
−53.094
3.468
12.237
1.00
50.43
C


ATOM
3734
CG
ARG
A
255
−52.678
3.736
13.696
1.00
50.82
C


ATOM
3737
CD
ARG
A
255
−52.242
5.211
13.887
1.00
51.32
C


ATOM
3740
NE
ARG
A
255
−51.003
5.522
13.155
1.00
51.81
N


ATOM
3742
CZ
ARG
A
255
−50.571
6.746
12.829
1.00
51.67
C


ATOM
3743
NH1
ARG
A
255
−51.264
7.839
13.146
1.00
51.63
N


ATOM
3746
NH2
ARG
A
255
−49.427
6.877
12.162
1.00
51.52
N


ATOM
3749
C
ARG
A
255
−54.160
2.040
10.497
1.00
50.32
C


ATOM
3750
O
ARG
A
255
−53.416
2.081
9.504
1.00
50.35
O


ATOM
3752
N
ASP
A
256
−55.492
2.005
10.412
1.00
50.17
N


ATOM
3753
CA
ASP
A
256
−56.190
2.090
9.128
1.00
50.18
C


ATOM
3755
CB
ASP
A
256
−57.259
1.009
9.037
1.00
50.37
C


ATOM
3758
CG
ASP
A
256
−58.095
1.130
7.780
1.00
50.96
C


ATOM
3759
OD1
ASP
A
256
−59.252
1.601
7.881
1.00
52.00
O


ATOM
3760
OD2
ASP
A
256
−57.581
.787
6.693
1.00
50.94
O


ATOM
3761
C
ASP
A
256
−56.838
3.470
8.945
1.00
49.85
C


ATOM
3762
O
ASP
A
256
−57.812
3.804
9.624
1.00
50.13
O


ATOM
3764
N
ARG
A
257
−56.315
4.263
8.013
1.00
49.11
N


ATOM
3765
CA
ARG
A
257
−56.712
5.665
7.913
1.00
48.59
C


ATOM
3767
CB
ARG
A
257
−55.622
6.553
8.529
1.00
48.79
C


ATOM
3770
CG
ARG
A
257
−55.318
6.275
10.006
1.00
49.85
C


ATOM
3773
CD
ARG
A
257
−56.538
6.480
10.908
1.00
51.01
C


ATOM
3776
NE
ARG
A
257
−56.198
7.274
12.089
1.00
51.80
N


ATOM
3778
CZ
ARG
A
257
−55.779
6.791
13.260
1.00
52.00
C


ATOM
3779
NH1
ARG
A
257
−55.643
5.486
13.468
1.00
51.64
N


ATOM
3782
NH2
ARG
A
257
−55.501
7.638
14.245
1.00
52.62
N


ATOM
3785
C
ARG
A
257
−56.988
6.112
6.477
1.00
47.58
C


ATOM
3786
O
ARG
A
257
−56.476
7.144
6.031
1.00
47.40
O


ATOM
3788
N
LEU
A
258
−57.814
5.352
5.761
1.00
46.18
N


ATOM
3789
CA
LEU
A
258
−58.094
5.672
4.358
1.00
44.78
C


ATOM
3791
CB
LEU
A
258
−58.777
4.508
3.629
1.00
44.57
C


ATOM
3794
CG
LEU
A
258
−58.917
4.731
2.121
1.00
43.79
C


ATOM
3796
CD1
LEU
A
258
−57.557
4.661
1.478
1.00
43.89
C


ATOM
3800
CD2
LEU
A
258
−59.850
3.733
1.492
1.00
42.88
C


ATOM
3804
C
LEU
A
258
−58.970
6.907
4.259
1.00
43.52
C


ATOM
3805
O
LEU
A
258
−58.643
7.849
3.545
1.00
43.42
O


ATOM
3807
N
ILE
A
259
−60.080
6.895
4.983
1.00
41.98
N


ATOM
3808
CA
ILE
A
259
−61.072
7.948
4.843
1.00
40.89
C


ATOM
3810
CB
ILE
A
259
−62.349
7.703
5.693
1.00
41.02
C


ATOM
3812
CG1
ILE
A
259
−62.846
6.248
5.570
1.00
41.82
C


ATOM
3815
CD1
ILE
A
259
−64.029
5.872
6.508
1.00
42.29
C


ATOM
3819
CG2
ILE
A
259
−63.446
8.662
5.262
1.00
40.47
C


ATOM
3823
C
ILE
A
259
−60.424
9.253
5.273
1.00
39.74
C


ATOM
3824
O
ILE
A
259
−60.607
10.291
4.635
1.00
39.25
O


ATOM
3826
N
GLU
A
260
−59.650
9.191
6.355
1.00
38.46
N


ATOM
3827
CA
GLU
A
260
−58.951
10.366
6.846
1.00
37.44
C


ATOM
3829
CB
GLU
A
260
−58.183
10.072
8.140
1.00
37.74
C


ATOM
3832
CG
GLU
A
260
−59.041
9.934
9.393
1.00
38.71
C


ATOM
3835
CD
GLU
A
260
−59.469
8.501
9.693
1.00
41.00
C


ATOM
3836
OE1
GLU
A
260
−59.273
7.594
8.842
1.00
42.70
O


ATOM
3837
OE2
GLU
A
260
−60.012
8.281
10.800
1.00
42.43
O


ATOM
3838
C
GLU
A
260
−57.995
10.841
5.764
1.00
35.94
C


ATOM
3839
O
GLU
A
260
−57.981
12.020
5.418
1.00
36.10
O


ATOM
3841
N
SER
A
261
−57.222
9.908
5.212
1.00
34.02
N


ATOM
3842
CA
SER
A
261
−56.263
10.229
4.152
1.00
32.47
C


ATOM
3844
CB
SER
A
261
−55.407
9.015
3.774
1.00
32.51
C


ATOM
3847
OG
SER
A
261
−54.253
8.940
4.584
1.00
33.14
O


ATOM
3849
C
SER
A
261
−56.894
10.764
2.889
1.00
30.82
C


ATOM
3850
O
SER
A
261
−56.199
11.306
2.062
1.00
30.79
O


ATOM
3852
N
PHE
A
262
−58.194
10.589
2.713
1.00
29.25
N


ATOM
3853
CA
PHE
A
262
−58.861
11.091
1.514
1.00
27.98
C


ATOM
3855
CB
PHE
A
262
−60.011
10.185
1.119
1.00
27.57
C


ATOM
3858
CG
PHE
A
262
−60.473
10.423
−.251
1.00
26.11
C


ATOM
3859
CD1
PHE
A
262
−59.763
9.914
−1.318
1.00
25.14
C


ATOM
3861
CE1
PHE
A
262
−60.169
10.147
−2.605
1.00
24.32
C


ATOM
3863
CZ
PHE
A
262
−61.284
10.917
−2.840
1.00
24.36
C


ATOM
3865
CE2
PHE
A
262
−61.987
11.445
−1.785
1.00
25.23
C


ATOM
3867
CD2
PHE
A
262
−61.575
11.201
−.492
1.00
25.61
C


ATOM
3869
C
PHE
A
262
−59.394
12.507
1.707
1.00
27.29
C


ATOM
3870
O
PHE
A
262
−59.275
13.359
.821
1.00
27.39
O


ATOM
3872
N
TYR
A
263
−60.025
12.723
2.856
1.00
26.19
N


ATOM
3873
CA
TYR
A
263
−60.415
14.052
3.310
1.00
25.22
C


ATOM
3875
CB
TYR
A
263
−60.975
13.942
4.735
1.00
25.21
C


ATOM
3878
CG
TYR
A
263
−61.037
15.199
5.578
1.00
25.37
C


ATOM
3879
CD1
TYR
A
263
−62.020
16.152
5.385
1.00
24.95
C


ATOM
3881
CE1
TYR
A
263
−62.084
17.282
6.197
1.00
26.54
C


ATOM
3883
CZ
TYR
A
263
−61.158
17.455
7.229
1.00
27.44
C


ATOM
3884
OH
TYR
A
263
−61.186
18.567
8.059
1.00
28.97
O


ATOM
3886
CE2
TYR
A
263
−60.185
16.512
7.438
1.00
27.10
C


ATOM
3888
CD2
TYR
A
263
−60.136
15.390
6.626
1.00
26.91
C


ATOM
3890
C
TYR
A
263
−59.190
14.946
3.238
1.00
24.29
C


ATOM
3891
O
TYR
A
263
−59.267
16.077
2.757
1.00
24.13
O


ATOM
3893
N
TRP
A
264
−58.055
14.415
3.682
1.00
23.01
N


ATOM
3894
CA
TRP
A
264
−56.789
15.121
3.567
1.00
22.32
C


ATOM
3896
CB
TRP
A
264
−55.642
14.261
4.119
1.00
22.26
C


ATOM
3899
CG
TRP
A
264
−54.326
14.874
3.860
1.00
22.16
C


ATOM
3900
CD1
TRP
A
264
−53.614
14.821
2.699
1.00
22.68
C


ATOM
3902
NE1
TRP
A
264
−52.456
15.538
2.818
1.00
23.08
N


ATOM
3904
CE2
TRP
A
264
−52.407
16.083
4.072
1.00
23.07
C


ATOM
3905
CD2
TRP
A
264
−53.579
15.686
4.753
1.00
22.18
C


ATOM
3906
CE3
TRP
A
264
−53.782
16.119
6.066
1.00
22.57
C


ATOM
3908
CZ3
TRP
A
264
−52.824
16.925
6.657
1.00
23.43
C


ATOM
3910
CH2
TRP
A
264
−51.656
17.297
5.955
1.00
24.14
C


ATOM
3912
CZ2
TRP
A
264
−51.433
16.886
4.663
1.00
23.64
C


ATOM
3914
C
TRP
A
264
−56.485
15.523
2.112
1.00
21.50
C


ATOM
3915
O
TRP
A
264
−56.179
16.682
1.820
1.00
21.29
O


ATOM
3917
N
ALA
A
265
−56.561
14.547
1.213
1.00
20.41
N


ATOM
3918
CA
ALA
A
265
−56.242
14.760
−.187
1.00
19.63
C


ATOM
3920
CB
ALA
A
265
−56.298
13.437
−.953
1.00
19.39
C


ATOM
3924
C
ALA
A
265
−57.176
15.792
−.798
1.00
19.07
C


ATOM
3925
O
ALA
A
265
−56.760
16.573
−1.633
1.00
18.99
O


ATOM
3927
N
VAL
A
266
−58.431
15.814
−.359
1.00
18.78
N


ATOM
3928
CA
VAL
A
266
−59.396
16.828
−.818
1.00
18.56
C


ATOM
3930
CB
VAL
A
266
−60.839
16.541
−.278
1.00
18.41
C


ATOM
3932
CG1
VAL
A
266
−61.405
15.298
−.935
1.00
18.88
C


ATOM
3936
CG2
VAL
A
266
−61.780
17.709
−.505
1.00
17.21
C


ATOM
3940
C
VAL
A
266
−58.947
18.257
−.450
1.00
18.43
C


ATOM
3941
O
VAL
A
266
−59.250
19.200
−1.171
1.00
18.83
O


ATOM
3943
N
GLY
A
267
−58.230
18.411
.663
1.00
17.96
N


ATOM
3944
CA
GLY
A
267
−57.714
19.712
1.067
1.00
17.45
C


ATOM
3947
C
GLY
A
267
−56.687
20.180
.070
1.00
17.20
C


ATOM
3948
O
GLY
A
267
−56.574
21.365
−.215
1.00
17.41
O


ATOM
3950
N
VAL
A
268
−55.959
19.219
−.480
1.00
16.88
N


ATOM
3951
CA
VAL
A
268
−54.822
19.487
−1.319
1.00
16.62
C


ATOM
3953
CB
VAL
A
268
−53.833
18.300
−1.290
1.00
16.19
C


ATOM
3955
CG1
VAL
A
268
−52.691
18.536
−2.240
1.00
15.50
C


ATOM
3959
CG2
VAL
A
268
−53.310
18.121
.103
1.00
15.30
C


ATOM
3963
C
VAL
A
268
−55.262
19.788
−2.736
1.00
17.15
C


ATOM
3964
O
VAL
A
268
−54.748
20.710
−3.350
1.00
17.12
O


ATOM
3966
N
ALA
A
269
−56.217
19.014
−3.243
1.00
17.99
N


ATOM
3967
CA
ALA
A
269
−56.688
19.158
−4.622
1.00
18.79
C


ATOM
3969
CB
ALA
A
269
−55.958
18.183
−5.551
1.00
18.26
C


ATOM
3973
C
ALA
A
269
−58.196
18.938
−4.672
1.00
19.70
C


ATOM
3974
O
ALA
A
269
−58.665
17.834
−4.958
1.00
19.50
O


ATOM
3976
N
PHE
A
270
−58.945
20.011
−4.417
1.00
21.09
N


ATOM
3977
CA
PHE
A
270
−60.393
19.910
−4.211
1.00
22.32
C


ATOM
3979
CB
PHE
A
270
−60.925
21.108
−3.404
1.00
22.49
C


ATOM
3982
CG
PHE
A
270
−61.193
22.321
−4.246
1.00
23.31
C


ATOM
3983
CD1
PHE
A
270
−62.423
22.483
−4.883
1.00
24.16
C


ATOM
3985
CE1
PHE
A
270
−62.662
23.569
−5.683
1.00
23.84
C


ATOM
3987
CZ
PHE
A
270
−61.675
24.497
−5.871
1.00
23.93
C


ATOM
3989
CE2
PHE
A
270
−60.448
24.349
−5.242
1.00
23.33
C


ATOM
3991
CD2
PHE
A
270
−60.213
23.268
−4.443
1.00
23.11
C


ATOM
3993
C
PHE
A
270
−61.173
19.820
−5.515
1.00
23.21
C


ATOM
3994
O
PHE
A
270
−62.142
19.069
−5.599
1.00
23.48
O


ATOM
3996
N
GLU
A
271
−60.771
20.599
−6.522
1.00
24.16
N


ATOM
3997
CA
GLU
A
271
−61.595
20.744
−7.741
1.00
25.01
C


ATOM
3999
CB
GLU
A
271
−61.065
21.863
−8.655
1.00
25.19
C


ATOM
4002
CG
GLU
A
271
−59.563
21.909
−8.794
1.00
27.45
C


ATOM
4005
CD
GLU
A
271
−58.857
22.910
−7.857
1.00
30.03
C


ATOM
4006
OE1
GLU
A
271
−58.905
24.145
−8.147
1.00
29.41
O


ATOM
4007
OE2
GLU
A
271
−58.227
22.431
−6.865
1.00
30.71
O


ATOM
4008
C
GLU
A
271
−61.799
19.391
−8.484
1.00
24.95
C


ATOM
4009
O
GLU
A
271
−60.972
18.480
−8.352
1.00
25.70
O


ATOM
4011
N
PRO
A
272
−62.918
19.242
−9.224
1.00
24.59
N


ATOM
4012
CA
PRO
A
272
−63.399
17.915
−9.656
1.00
24.38
C


ATOM
4014
CB
PRO
A
272
−64.675
18.243
−10.430
1.00
24.25
C


ATOM
4017
CG
PRO
A
272
−65.105
19.536
−9.869
1.00
24.53
C


ATOM
4020
CD
PRO
A
272
−63.855
20.295
−9.639
1.00
24.42
C


ATOM
4023
C
PRO
A
272
−62.475
17.111
−10.549
1.00
24.29
C


ATOM
4024
O
PRO
A
272
−62.406
15.886
−10.410
1.00
24.45
O


ATOM
4025
N
GLN
A
273
−61.777
17.781
−11.462
1.00
24.13
N


ATOM
4026
CA
GLN
A
273
−60.998
17.064
−12.461
1.00
24.13
C


ATOM
4028
CB
GLN
A
273
−60.523
17.983
−13.567
1.00
23.84
C


ATOM
4031
CG
GLN
A
273
−59.556
19.041
−13.117
1.00
24.37
C


ATOM
4034
CD
GLN
A
273
−60.222
20.360
−12.775
1.00
25.22
C


ATOM
4035
OE1
GLN
A
273
−61.392
20.412
−12.367
1.00
25.71
O


ATOM
4036
NE2
GLN
A
273
−59.468
21.441
−12.932
1.00
24.95
N


ATOM
4039
C
GLN
A
273
−59.808
16.327
−11.877
1.00
24.33
C


ATOM
4040
O
GLN
A
273
−59.224
15.486
−12.555
1.00
24.91
O


ATOM
4042
N
TYR
A
274
−59.462
16.607
−10.626
1.00
24.27
N


ATOM
4043
CA
TYR
A
274
−58.211
16.119
−10.067
1.00
24.38
C


ATOM
4045
CB
TYR
A
274
−57.639
17.180
−9.138
1.00
24.36
C


ATOM
4048
CG
TYR
A
274
−57.066
18.398
−9.819
1.00
24.35
C


ATOM
4049
CD1
TYR
A
274
−56.249
18.291
−10.935
1.00
24.13
C


ATOM
4051
CE1
TYR
A
274
−55.716
19.409
−11.542
1.00
23.75
C


ATOM
4053
CZ
TYR
A
274
−55.968
20.646
−11.019
1.00
24.14
C


ATOM
4054
OH
TYR
A
274
−55.431
21.762
−11.597
1.00
24.45
O


ATOM
4056
CE2
TYR
A
274
−56.755
20.780
−9.903
1.00
24.92
C


ATOM
4058
CD2
TYR
A
274
−57.297
19.658
−9.308
1.00
24.74
C


ATOM
4060
C
TYR
A
274
−58.319
14.796
−9.310
1.00
24.63
C


ATOM
4061
O
TYR
A
274
−57.681
14.613
−8.276
1.00
24.47
O


ATOM
4063
N
SER
A
275
−59.097
13.852
−9.821
1.00
25.00
N


ATOM
4064
CA
SER
A
275
−59.254
12.575
−9.116
1.00
25.11
C


ATOM
4066
CB
SER
A
275
−60.244
11.664
−9.842
1.00
25.14
C


ATOM
4069
OG
SER
A
275
−61.537
12.252
−9.852
1.00
25.96
O


ATOM
4071
C
SER
A
275
−57.907
11.885
−8.963
1.00
24.94
C


ATOM
4072
O
SER
A
275
−57.544
11.439
−7.877
1.00
24.57
O


ATOM
4074
N
ASP
A
276
−57.149
11.822
−10.052
1.00
25.01
N


ATOM
4075
CA
ASP
A
276
−55.840
11.187
−9.990
1.00
24.92
C


ATOM
4077
CB
ASP
A
276
−55.102
11.257
−11.324
1.00
25.02
C


ATOM
4080
CG
ASP
A
276
−55.827
10.495
−12.407
1.00
26.27
C


ATOM
4081
OD1
ASP
A
276
−56.372
9.411
−12.080
1.00
26.65
O


ATOM
4082
OD2
ASP
A
276
−55.880
10.991
−13.563
1.00
28.21
O


ATOM
4083
C
ASP
A
276
−55.026
11.798
−8.881
1.00
24.30
C


ATOM
4084
O
ASP
A
276
−54.454
11.064
−8.099
1.00
24.64
O


ATOM
4086
N
CYS
A
277
−55.005
13.124
−8.772
1.00
23.61
N


ATOM
4087
CA
CYS
A
277
−54.210
13.746
−7.715
1.00
22.91
C


ATOM
4089
CB
CYS
A
277
−54.262
15.268
−7.748
1.00
22.83
C


ATOM
4092
SG
CYS
A
277
−53.048
15.994
−6.615
1.00
22.43
S


ATOM
4094
C
CYS
A
277
−54.653
13.251
−6.357
1.00
22.39
C


ATOM
4095
O
CYS
A
277
−53.822
12.833
−5.559
1.00
22.23
O


ATOM
4097
N
ARG
A
278
−55.962
13.262
−6.115
1.00
22.01
N


ATOM
4098
CA
ARG
A
278
−56.519
12.813
−4.829
1.00
21.40
C


ATOM
4100
CB
ARG
A
278
−58.021
13.053
−4.753
1.00
21.07
C


ATOM
4103
CG
ARG
A
278
−58.382
14.509
−4.754
1.00
20.39
C


ATOM
4106
CD
ARG
A
278
−59.852
14.698
−4.527
1.00
20.05
C


ATOM
4109
NE
ARG
A
278
−60.683
14.331
−5.675
1.00
18.86
N


ATOM
4111
CZ
ARG
A
278
−60.993
15.144
−6.679
1.00
18.58
C


ATOM
4112
NH1
ARG
A
278
−60.530
16.391
−6.730
1.00
18.45
N


ATOM
4115
NH2
ARG
A
278
−61.775
14.702
−7.647
1.00
19.04
N


ATOM
4118
C
ARG
A
278
−56.224
11.358
−4.530
1.00
21.12
C


ATOM
4119
O
ARG
A
278
−55.804
11.050
−3.434
1.00
21.59
O


ATOM
4121
N
ASN
A
279
−56.412
10.467
−5.493
1.00
20.94
N


ATOM
4122
CA
ASN
A
279
−56.168
9.044
−5.243
1.00
21.08
C


ATOM
4124
CB
ASN
A
279
−56.672
8.182
−6.401
1.00
21.54
C


ATOM
4127
CG
ASN
A
279
−58.199
8.346
−6.642
1.00
23.94
C


ATOM
4128
OD1
ASN
A
279
−58.920
8.941
−5.823
1.00
26.47
O


ATOM
4129
ND2
ASN
A
279
−58.685
7.825
−7.769
1.00
25.52
N


ATOM
4132
C
ASN
A
279
−54.699
8.798
−4.972
1.00
20.30
C


ATOM
4133
O
ASN
A
279
−54.337
8.200
−3.977
1.00
19.95
O


ATOM
4135
N
SER
A
280
−53.851
9.297
−5.856
1.00
20.02
N


ATOM
4136
CA
SER
A
280
−52.409
9.335
−5.612
1.00
19.32
C


ATOM
4138
CB
SER
A
280
−51.725
10.304
−6.580
1.00
19.35
C


ATOM
4141
OG
SER
A
280
−50.624
9.705
−7.212
1.00
19.77
O


ATOM
4143
C
SER
A
280
−52.122
9.745
−4.166
1.00
18.55
C


ATOM
4144
O
SER
A
280
−51.514
8.976
−3.436
1.00
18.98
O


ATOM
4146
N
VAL
A
281
−52.580
10.923
−3.743
1.00
17.45
N


ATOM
4147
CA
VAL
A
281
−52.235
11.444
−2.416
1.00
17.01
C


ATOM
4149
CB
VAL
A
281
−52.650
12.934
−2.235
1.00
17.20
C


ATOM
4151
CG1
VAL
A
281
−52.492
13.376
−.787
1.00
16.76
C


ATOM
4155
CG2
VAL
A
281
−51.826
13.847
−3.128
1.00
17.13
C


ATOM
4159
C
VAL
A
281
−52.861
10.607
−1.301
1.00
16.82
C


ATOM
4160
O
VAL
A
281
−52.217
10.314
−.289
1.00
16.81
O


ATOM
4162
N
ALA
A
282
−54.118
10.225
−1.481
1.00
16.55
N


ATOM
4163
CA
ALA
A
282
−54.774
9.304
−.559
1.00
16.31
C


ATOM
4165
CB
ALA
A
282
−56.174
8.951
−1.060
1.00
16.08
C


ATOM
4169
C
ALA
A
282
−53.946
8.036
−.355
1.00
16.31
C


ATOM
4170
O
ALA
A
282
−53.671
7.653
.762
1.00
16.16
O


ATOM
4172
N
LYS
A
283
−53.539
7.395
−1.443
1.00
16.92
N


ATOM
4173
CA
LYS
A
283
−52.773
6.139
−1.368
1.00
17.39
C


ATOM
4175
CB
LYS
A
283
−52.454
5.596
−2.773
1.00
17.31
C


ATOM
4178
CG
LYS
A
283
−53.680
5.066
−3.519
1.00
17.85
C


ATOM
4181
CD
LYS
A
283
−53.389
4.527
−4.941
1.00
18.98
C


ATOM
4184
CE
LYS
A
283
−54.709
4.275
−5.718
1.00
19.79
C


ATOM
4187
NZ
LYS
A
283
−54.695
3.091
−6.625
1.00
20.11
N


ATOM
4191
C
LYS
A
283
−51.483
6.338
−.591
1.00
17.82
C


ATOM
4192
O
LYS
A
283
−51.170
5.577
.331
1.00
17.37
O


ATOM
4194
N
MET
A
284
−50.744
7.381
−.964
1.00
18.49
N


ATOM
4195
CA
MET
A
284
−49.434
7.622
−.373
1.00
18.82
C


ATOM
4197
CB
MET
A
284
−48.688
8.795
−1.047
1.00
18.84
C


ATOM
4200
CG
MET
A
284
−48.118
8.506
−2.444
1.00
18.47
C


ATOM
4203
SD
MET
A
284
−47.540
6.819
−2.718
1.00
19.50
S


ATOM
4204
CE
MET
A
284
−49.088
5.982
−3.094
1.00
19.36
C


ATOM
4208
C
MET
A
284
−49.575
7.848
1.115
1.00
18.95
C


ATOM
4209
O
MET
A
284
−48.901
7.166
1.892
1.00
19.29
O


ATOM
4211
N
PHE
A
285
−50.465
8.762
1.507
1.00
18.94
N


ATOM
4212
CA
PHE
A
285
−50.671
9.082
2.930
1.00
19.26
C


ATOM
4214
CB
PHE
A
285
−51.714
10.185
3.073
1.00
19.72
C


ATOM
4217
CG
PHE
A
285
−51.575
11.016
4.328
1.00
21.97
C


ATOM
4218
CD1
PHE
A
285
−50.335
11.202
4.954
1.00
24.49
C


ATOM
4220
CE1
PHE
A
285
−50.209
11.996
6.086
1.00
25.11
C


ATOM
4222
CZ
PHE
A
285
−51.321
12.634
6.597
1.00
26.40
C


ATOM
4224
CE2
PHE
A
285
−52.566
12.470
5.977
1.00
26.44
C


ATOM
4226
CD2
PHE
A
285
−52.678
11.670
4.846
1.00
24.55
C


ATOM
4228
C
PHE
A
285
−51.097
7.879
3.770
1.00
18.74
C


ATOM
4229
O
PHE
A
285
−50.700
7.729
4.924
1.00
18.31
O


ATOM
4231
N
SER
A
286
−51.903
7.019
3.169
1.00
18.49
N


ATOM
4232
CA
SER
A
286
−52.281
5.772
3.791
1.00
18.38
C


ATOM
4234
CB
SER
A
286
−53.302
5.065
2.926
1.00
18.45
C


ATOM
4237
OG
SER
A
286
−54.389
5.949
2.687
1.00
19.82
O


ATOM
4239
C
SER
A
286
−51.070
4.888
4.048
1.00
18.08
C


ATOM
4240
O
SER
A
286
−50.931
4.376
5.146
1.00
18.18
O


ATOM
4242
N
PHE
A
287
−50.195
4.714
3.054
1.00
17.72
N


ATOM
4243
CA
PHE
A
287
−48.911
4.024
3.279
1.00
17.23
C


ATOM
4245
CB
PHE
A
287
−48.133
3.801
1.977
1.00
17.09
C


ATOM
4248
CG
PHE
A
287
−48.513
2.555
1.258
1.00
17.26
C


ATOM
4249
CD1
PHE
A
287
−48.220
1.315
1.802
1.00
18.31
C


ATOM
4251
CE1
PHE
A
287
−48.583
.129
1.131
1.00
18.47
C


ATOM
4253
CZ
PHE
A
287
−49.237
.198
−.091
1.00
17.88
C


ATOM
4255
CE2
PHE
A
287
−49.530
1.441
−.642
1.00
17.57
C


ATOM
4257
CD2
PHE
A
287
−49.168
2.608
.031
1.00
17.79
C


ATOM
4259
C
PHE
A
287
−48.022
4.787
4.277
1.00
16.92
C


ATOM
4260
O
PHE
A
287
−47.369
4.173
5.143
1.00
17.18
O


ATOM
4262
N
VAL
A
288
−47.989
6.113
4.174
1.00
16.00
N


ATOM
4263
CA
VAL
A
288
−47.148
6.883
5.080
1.00
15.45
C


ATOM
4265
CB
VAL
A
288
−47.289
8.399
4.863
1.00
15.21
C


ATOM
4267
CG1
VAL
A
288
−46.433
9.162
5.852
1.00
14.24
C


ATOM
4271
CG2
VAL
A
288
−46.898
8.757
3.450
1.00
14.46
C


ATOM
4275
C
VAL
A
288
−47.479
6.499
6.527
1.00
15.46
C


ATOM
4276
O
VAL
A
288
−46.590
6.160
7.299
1.00
14.79
O


ATOM
4278
N
THR
A
289
−48.759
6.494
6.880
1.00
15.62
N


ATOM
4279
CA
THR
A
289
−49.110
6.272
8.274
1.00
16.09
C


ATOM
4281
CB
THR
A
289
−50.602
6.557
8.588
1.00
15.72
C


ATOM
4283
OG1
THR
A
289
−51.422
5.827
7.708
1.00
16.52
O


ATOM
4285
CG2
THR
A
289
−50.927
8.007
8.371
1.00
16.56
C


ATOM
4289
C
THR
A
289
−48.677
4.879
8.727
1.00
16.34
C


ATOM
4290
O
THR
A
289
−48.234
4.707
9.881
1.00
16.68
O


ATOM
4292
N
ILE
A
290
−48.737
3.904
7.820
1.00
16.29
N


ATOM
4293
CA
ILE
A
290
−48.347
2.538
8.166
1.00
16.56
C


ATOM
4295
CB
ILE
A
290
−48.734
1.516
7.096
1.00
16.48
C


ATOM
4297
CG1
ILE
A
290
−50.247
1.440
6.944
1.00
16.91
C


ATOM
4300
CD1
ILE
A
290
−50.677
.564
5.811
1.00
16.52
C


ATOM
4304
CG2
ILE
A
290
−48.262
.145
7.488
1.00
16.52
C


ATOM
4308
C
ILE
A
290
−46.842
2.456
8.402
1.00
16.94
C


ATOM
4309
O
ILE
A
290
−46.404
1.968
9.443
1.00
17.06
O


ATOM
4311
N
ILE
A
291
−46.048
2.942
7.451
1.00
17.22
N


ATOM
4312
CA
ILE
A
291
−44.595
2.969
7.640
1.00
17.20
C


ATOM
4314
CB
ILE
A
291
−43.842
3.534
6.405
1.00
17.32
C


ATOM
4316
CG1
ILE
A
291
−44.172
2.744
5.125
1.00
17.61
C


ATOM
4319
CD1
ILE
A
291
−44.004
1.265
5.250
1.00
18.21
C


ATOM
4323
CG2
ILE
A
291
−42.336
3.550
6.637
1.00
16.64
C


ATOM
4327
C
ILE
A
291
−44.262
3.794
8.886
1.00
17.36
C


ATOM
4328
O
ILE
A
291
−43.420
3.413
9.670
1.00
17.06
O


ATOM
4330
N
ASP
A
292
−44.950
4.906
9.093
1.00
18.12
N


ATOM
4331
CA
ASP
A
292
−44.627
5.755
10.227
1.00
18.83
C


ATOM
4333
CB
ASP
A
292
−45.511
7.017
10.285
1.00
19.22
C


ATOM
4336
CG
ASP
A
292
−45.185
7.920
11.485
1.00
19.93
C


ATOM
4337
OD1
ASP
A
292
−44.062
8.460
11.544
1.00
21.80
O


ATOM
4338
OD2
ASP
A
292
−46.048
8.080
12.375
1.00
20.46
O


ATOM
4339
C
ASP
A
292
−44.745
4.947
11.509
1.00
18.96
C


ATOM
4340
O
ASP
A
292
−43.902
5.094
12.394
1.00
19.05
O


ATOM
4342
N
ASP
A
293
−45.774
4.097
11.610
1.00
19.05
N


ATOM
4343
CA
ASP
A
293
−45.951
3.273
12.817
1.00
18.99
C


ATOM
4345
CB
ASP
A
293
−47.237
2.443
12.790
1.00
18.95
C


ATOM
4348
CG
ASP
A
293
−48.496
3.272
12.921
1.00
19.27
C


ATOM
4349
OD1
ASP
A
293
−48.483
4.361
13.523
1.00
21.51
O


ATOM
4350
OD2
ASP
A
293
−49.536
2.812
12.421
1.00
19.61
O


ATOM
4351
C
ASP
A
293
−44.783
2.322
12.935
1.00
19.07
C


ATOM
4352
O
ASP
A
293
−44.244
2.135
14.024
1.00
19.35
O


ATOM
4354
N
ILE
A
294
−44.390
1.730
11.805
1.00
19.10
N


ATOM
4355
CA
ILE
A
294
−43.320
.727
11.786
1.00
19.04
C


ATOM
4357
CB
ILE
A
294
−43.137
.109
10.386
1.00
18.49
C


ATOM
4359
CG1
ILE
A
294
−44.343
−.754
10.038
1.00
17.60
C


ATOM
4362
CD1
ILE
A
294
−44.204
−1.504
8.752
1.00
16.77
C


ATOM
4366
CG2
ILE
A
294
−41.895
−.745
10.348
1.00
18.77
C


ATOM
4370
C
ILE
A
294
−41.974
1.273
12.298
1.00
19.55
C


ATOM
4371
O
ILE
A
294
−41.234
.550
12.974
1.00
19.75
O


ATOM
4373
N
TYR
A
295
−41.649
2.525
11.983
1.00
19.75
N


ATOM
4374
CA
TYR
A
295
−40.411
3.103
12.476
1.00
19.91
C


ATOM
4376
CB
TYR
A
295
−39.856
4.178
11.542
1.00
19.66
C


ATOM
4379
CG
TYR
A
295
−39.206
3.695
10.245
1.00
18.19
C


ATOM
4380
CD1
TYR
A
295
−37.821
3.727
10.075
1.00
16.19
C


ATOM
4382
CE1
TYR
A
295
−37.222
3.331
8.882
1.00
14.68
C


ATOM
4384
CZ
TYR
A
295
−38.004
2.906
7.828
1.00
15.04
C


ATOM
4385
OH
TYR
A
295
−37.428
2.485
6.631
1.00
12.25
O


ATOM
4387
CE2
TYR
A
295
−39.383
2.871
7.975
1.00
16.48
C


ATOM
4389
CD2
TYR
A
295
−39.975
3.271
9.172
1.00
17.01
C


ATOM
4391
C
TYR
A
295
−40.647
3.677
13.858
1.00
20.73
C


ATOM
4392
O
TYR
A
295
−39.749
3.635
14.717
1.00
21.64
O


ATOM
4394
N
ASP
A
296
−41.843
4.200
14.103
1.00
21.38
N


ATOM
4395
CA
ASP
A
296
−42.100
4.865
15.391
1.00
22.20
C


ATOM
4397
CB
ASP
A
296
−43.421
5.644
15.391
1.00
22.69
C


ATOM
4400
CG
ASP
A
296
−43.567
6.548
16.607
1.00
23.85
C


ATOM
4401
OD1
ASP
A
296
−42.568
7.179
17.014
1.00
25.49
O


ATOM
4402
OD2
ASP
A
296
−44.689
6.648
17.145
1.00
26.12
O


ATOM
4403
C
ASP
A
296
−42.106
3.890
16.549
1.00
22.10
C


ATOM
4404
O
ASP
A
296
−41.376
4.082
17.506
1.00
22.14
O


ATOM
4406
N
VAL
A
297
−42.900
2.828
16.435
1.00
22.25
N


ATOM
4407
CA
VAL
A
297
−43.163
1.937
17.570
1.00
22.22
C


ATOM
4409
CB
VAL
A
297
−44.635
2.116
18.057
1.00
22.07
C


ATOM
4411
CG1
VAL
A
297
−44.958
3.587
18.215
1.00
21.63
C


ATOM
4415
CG2
VAL
A
297
−45.618
1.459
17.099
1.00
20.76
C


ATOM
4419
C
VAL
A
297
−42.861
.422
17.373
1.00
22.49
C


ATOM
4420
O
VAL
A
297
−42.517
−.255
18.343
1.00
22.40
O


ATOM
4422
N
TYR
A
298
−42.990
−.117
16.157
1.00
22.69
N


ATOM
4423
CA
TYR
A
298
−43.065
−1.579
15.993
1.00
22.90
C


ATOM
4425
CB
TYR
A
298
−44.089
−1.974
14.934
1.00
22.77
C


ATOM
4428
CG
TYR
A
298
−44.341
−3.469
14.932
1.00
23.74
C


ATOM
4429
CD1
TYR
A
298
−45.249
−4.043
15.819
1.00
25.72
C


ATOM
4431
CE1
TYR
A
298
−45.484
−5.435
15.833
1.00
26.12
C


ATOM
4433
CZ
TYR
A
298
−44.798
−6.247
14.954
1.00
25.59
C


ATOM
4434
OH
TYR
A
298
−45.027
−7.593
14.970
1.00
25.33
O


ATOM
4436
CE2
TYR
A
298
−43.890
−5.702
14.062
1.00
24.79
C


ATOM
4438
CD2
TYR
A
298
−43.661
−4.317
14.061
1.00
24.20
C


ATOM
4440
C
TYR
A
298
−41.750
−2.258
15.649
1.00
23.02
C


ATOM
4441
O
TYR
A
298
−41.387
−3.257
16.256
1.00
22.80
O


ATOM
4443
N
GLY
A
299
−41.069
−1.747
14.634
1.00
23.47
N


ATOM
4444
CA
GLY
A
299
−39.839
−2.361
14.149
1.00
23.36
C


ATOM
4447
C
GLY
A
299
−38.631
−1.978
14.984
1.00
23.27
C


ATOM
4448
O
GLY
A
299
−38.532
−.858
15.501
1.00
23.35
O


ATOM
4450
N
THR
A
300
−37.702
−2.918
15.098
1.00
23.11
N


ATOM
4451
CA
THR
A
300
−36.459
−2.682
15.797
1.00
22.90
C


ATOM
4453
CB
THR
A
300
−35.869
−3.957
16.386
1.00
22.83
C


ATOM
4455
OG1
THR
A
300
−35.328
−4.756
15.328
1.00
22.61
O


ATOM
4457
CG2
THR
A
300
−36.928
−4.732
17.162
1.00
22.18
C


ATOM
4461
C
THR
A
300
−35.482
−2.120
14.796
1.00
23.00
C


ATOM
4462
O
THR
A
300
−35.606
−2.364
13.602
1.00
23.13
O


ATOM
4464
N
LEU
A
301
−34.496
−1.394
15.309
1.00
23.07
N


ATOM
4465
CA
LEU
A
301
−33.596
−.572
14.498
1.00
22.84
C


ATOM
4467
CB
LEU
A
301
−32.515
.007
15.403
1.00
22.80
C


ATOM
4470
CG
LEU
A
301
−31.965
1.412
15.182
1.00
22.39
C


ATOM
4472
CD1
LEU
A
301
−32.983
2.365
14.587
1.00
22.44
C


ATOM
4476
CD2
LEU
A
301
−31.482
1.919
16.536
1.00
22.19
C


ATOM
4480
C
LEU
A
301
−32.964
−1.358
13.361
1.00
22.95
C


ATOM
4481
O
LEU
A
301
−32.910
−.888
12.232
1.00
22.50
O


ATOM
4483
N
ASP
A
302
−32.511
−2.569
13.673
1.00
23.35
N


ATOM
4484
CA
ASP
A
302
−31.929
−3.469
12.670
1.00
23.65
C


ATOM
4486
CB
ASP
A
302
−31.332
−4.747
13.328
1.00
24.12
C


ATOM
4489
CG
ASP
A
302
−29.984
−4.498
14.057
1.00
25.23
C


ATOM
4490
OD1
ASP
A
302
−29.003
−4.051
13.409
1.00
26.43
O


ATOM
4491
OD2
ASP
A
302
−29.896
−4.780
15.277
1.00
26.92
O


ATOM
4492
C
ASP
A
302
−32.945
−3.859
11.581
1.00
23.11
C


ATOM
4493
O
ASP
A
302
−32.544
−4.086
10.445
1.00
23.05
O


ATOM
4495
N
GLU
A
303
−34.235
−3.965
11.927
1.00
22.63
N


ATOM
4496
CA
GLU
A
303
−35.295
−4.248
10.931
1.00
22.40
C


ATOM
4498
CB
GLU
A
303
−36.625
−4.675
11.589
1.00
22.20
C


ATOM
4501
CG
GLU
A
303
−36.537
−5.956
12.426
1.00
22.66
C


ATOM
4504
CD
GLU
A
303
−37.832
−6.341
13.159
1.00
22.69
C


ATOM
4505
OE1
GLU
A
303
−38.614
−5.449
13.568
1.00
22.23
O


ATOM
4506
OE2
GLU
A
303
−38.051
−7.558
13.338
1.00
21.64
O


ATOM
4507
C
GLU
A
303
−35.533
−3.017
10.067
1.00
22.35
C


ATOM
4508
O
GLU
A
303
−35.658
−3.105
8.841
1.00
22.27
O


ATOM
4510
N
LEU
A
304
−35.597
−1.866
10.724
1.00
22.34
N


ATOM
4511
CA
LEU
A
304
−35.776
−.607
10.040
1.00
22.26
C


ATOM
4513
CB
LEU
A
304
−35.861
.537
11.052
1.00
22.49
C


ATOM
4516
CG
LEU
A
304
−37.089
.517
11.973
1.00
23.05
C


ATOM
4518
CD1
LEU
A
304
−37.109
1.708
12.932
1.00
23.46
C


ATOM
4522
CD2
LEU
A
304
−38.353
.511
11.142
1.00
23.96
C


ATOM
4526
C
LEU
A
304
−34.634
−.383
9.063
1.00
22.09
C


ATOM
4527
O
LEU
A
304
−34.873
.094
7.969
1.00
22.32
O


ATOM
4529
N
GLU
A
305
−33.408
−.740
9.450
1.00
21.91
N


ATOM
4530
CA
GLU
A
305
−32.259
−.685
8.541
1.00
21.93
C


ATOM
4532
CB
GLU
A
305
−30.988
−1.194
9.219
1.00
22.35
C


ATOM
4535
CG
GLU
A
305
−30.363
−.258
10.249
1.00
24.12
C


ATOM
4538
CD
GLU
A
305
−29.751
1.002
9.649
1.00
26.11
C


ATOM
4539
OE1
GLU
A
305
−29.275
1.846
10.454
1.00
25.68
O


ATOM
4540
OE2
GLU
A
305
−29.756
1.146
8.391
1.00
27.67
O


ATOM
4541
C
GLU
A
305
−32.475
−1.524
7.295
1.00
21.54
C


ATOM
4542
O
GLU
A
305
−32.010
−1.162
6.220
1.00
21.75
O


ATOM
4544
N
LEU
A
306
−33.155
−2.660
7.448
1.00
21.21
N


ATOM
4545
CA
LEU
A
306
−33.459
−3.557
6.316
1.00
20.66
C


ATOM
4547
CB
LEU
A
306
−33.880
−4.956
6.795
1.00
20.26
C


ATOM
4550
CG
LEU
A
306
−32.725
−5.854
7.220
1.00
19.57
C


ATOM
4552
CD1
LEU
A
306
−33.261
−7.150
7.777
1.00
20.18
C


ATOM
4556
CD2
LEU
A
306
−31.791
−6.120
6.061
1.00
18.29
C


ATOM
4560
C
LEU
A
306
−34.535
−2.967
5.416
1.00
20.23
C


ATOM
4561
O
LEU
A
306
−34.412
−2.966
4.197
1.00
19.97
O


ATOM
4563
N
PHE
A
307
−35.588
−2.457
6.023
1.00
19.99
N


ATOM
4564
CA
PHE
A
307
−36.635
−1.861
5.237
1.00
20.00
C


ATOM
4566
CB
PHE
A
307
−37.771
−1.388
6.118
1.00
20.08
C


ATOM
4569
CG
PHE
A
307
−39.011
−1.115
5.370
1.00
19.05
C


ATOM
4570
CD1
PHE
A
307
−39.895
−2.118
5.119
1.00
18.77
C


ATOM
4572
CE1
PHE
A
307
−41.037
−1.873
4.421
1.00
20.07
C


ATOM
4574
CZ
PHE
A
307
−41.296
−.621
3.959
1.00
19.63
C


ATOM
4576
CE2
PHE
A
307
−40.411
.391
4.199
1.00
19.27
C


ATOM
4578
CD2
PHE
A
307
−39.278
.142
4.900
1.00
19.17
C


ATOM
4580
C
PHE
A
307
−36.087
−.695
4.447
1.00
20.25
C


ATOM
4581
O
PHE
A
307
−36.356
−.589
3.242
1.00
20.08
O


ATOM
4583
N
THR
A
308
−35.325
.167
5.137
1.00
20.51
N


ATOM
4584
CA
THR
A
308
−34.737
1.379
4.538
1.00
20.60
C


ATOM
4586
CB
THR
A
308
−33.860
2.177
5.541
1.00
20.41
C


ATOM
4588
OG1
THR
A
308
−34.672
2.661
6.611
1.00
20.05
O


ATOM
4590
CG2
THR
A
308
−33.198
3.376
4.864
1.00
20.38
C


ATOM
4594
C
THR
A
308
−33.895
1.009
3.331
1.00
20.82
C


ATOM
4595
O
THR
A
308
−34.057
1.571
2.243
1.00
20.45
O


ATOM
4597
N
ASP
A
309
−33.020
.031
3.522
1.00
21.32
N


ATOM
4598
CA
ASP
A
309
−32.138
−.399
2.449
1.00
21.88
C


ATOM
4600
CB
ASP
A
309
−31.042
−1.324
2.971
1.00
22.44
C


ATOM
4603
CG
ASP
A
309
−30.164
−1.847
1.857
1.00
25.69
C


ATOM
4604
OD1
ASP
A
309
−29.599
−1.010
1.089
1.00
28.02
O


ATOM
4605
OD2
ASP
A
309
−30.076
−3.100
1.730
1.00
30.59
O


ATOM
4606
C
ASP
A
309
−32.928
−1.085
1.357
1.00
21.17
C


ATOM
4607
O
ASP
A
309
−32.601
−.948
.188
1.00
20.95
O


ATOM
4609
N
ALA
A
310
−33.965
−1.818
1.757
1.00
20.99
N


ATOM
4610
CA
ALA
A
310
−34.885
−2.474
.828
1.00
20.91
C


ATOM
4612
CB
ALA
A
310
−35.931
−3.244
1.591
1.00
20.77
C


ATOM
4616
C
ALA
A
310
−35.567
−1.501
−.123
1.00
21.08
C


ATOM
4617
O
ALA
A
310
−35.745
−1.816
−1.305
1.00
20.76
O


ATOM
4619
N
VAL
A
311
−35.958
−.334
.402
1.00
21.51
N


ATOM
4620
CA
VAL
A
311
−36.594
.719
−.396
1.00
21.50
C


ATOM
4622
CB
VAL
A
311
−37.424
1.715
.477
1.00
21.73
C


ATOM
4624
CG1
VAL
A
311
−38.724
1.076
.941
1.00
21.01
C


ATOM
4628
CG2
VAL
A
311
−37.765
2.993
−.292
1.00
21.88
C


ATOM
4632
C
VAL
A
311
−35.553
1.445
−1.230
1.00
21.67
C


ATOM
4633
O
VAL
A
311
−35.811
1.728
−2.389
1.00
21.71
O


ATOM
4635
N
GLU
A
312
−34.381
1.724
−.663
1.00
22.25
N


ATOM
4636
CA
GLU
A
312
−33.248
2.284
−1.443
1.00
23.00
C


ATOM
4638
CB
GLU
A
312
−31.951
2.331
−.608
1.00
23.36
C


ATOM
4641
CG
GLU
A
312
−31.897
3.383
.511
1.00
24.72
C


ATOM
4644
CD
GLU
A
312
−30.526
3.469
1.189
1.00
27.09
C


ATOM
4645
OE1
GLU
A
312
−30.083
4.608
1.458
1.00
30.07
O


ATOM
4646
OE2
GLU
A
312
−29.885
2.418
1.454
1.00
27.86
O


ATOM
4647
C
GLU
A
312
−32.954
1.498
−2.731
1.00
23.13
C


ATOM
4648
O
GLU
A
312
−32.851
2.064
−3.803
1.00
22.88
O


ATOM
4650
N
ARG
A
313
−32.819
.188
−2.615
1.00
23.74
N


ATOM
4651
CA
ARG
A
313
−32.400
−.628
−3.743
1.00
24.46
C


ATOM
4653
CB
ARG
A
313
−31.758
−1.916
−3.232
1.00
25.01
C


ATOM
4656
CG
ARG
A
313
−30.421
−1.667
−2.481
1.00
27.74
C


ATOM
4659
CD
ARG
A
313
−29.715
−2.963
−2.072
1.00
31.41
C


ATOM
4662
NE
ARG
A
313
−30.675
−3.921
−1.499
1.00
34.94
N


ATOM
4664
CZ
ARG
A
313
−31.203
−4.974
−2.138
1.00
37.87
C


ATOM
4665
NH1
ARG
A
313
−30.861
−5.282
−3.402
1.00
38.35
N


ATOM
4668
NH2
ARG
A
313
−32.079
−5.746
−1.495
1.00
38.82
N


ATOM
4671
C
ARG
A
313
−33.523
−.914
−4.739
1.00
24.27
C


ATOM
4672
O
ARG
A
313
−33.257
−1.178
−5.898
1.00
24.17
O


ATOM
4674
N
TRP
A
314
−34.770
−.873
−4.282
1.00
24.59
N


ATOM
4675
CA
TRP
A
314
−35.939
−1.042
−5.142
1.00
24.66
C


ATOM
4677
CB
TRP
A
314
−36.175
.234
−5.961
1.00
24.39
C


ATOM
4680
CG
TRP
A
314
−37.575
.382
−6.386
1.00
22.61
C


ATOM
4681
CD1
TRP
A
314
−38.073
.204
−7.635
1.00
21.66
C


ATOM
4683
NE1
TRP
A
314
−39.429
.403
−7.634
1.00
21.42
N


ATOM
4685
CE2
TRP
A
314
−39.829
.707
−6.360
1.00
20.99
C


ATOM
4686
CD2
TRP
A
314
−38.683
.701
−5.550
1.00
21.41
C


ATOM
4687
CE3
TRP
A
314
−38.817
.991
−4.191
1.00
21.48
C


ATOM
4689
CZ3
TRP
A
314
−40.080
1.277
−3.693
1.00
21.38
C


ATOM
4691
CH2
TRP
A
314
−41.204
1.269
−4.523
1.00
20.84
C


ATOM
4693
CZ2
TRP
A
314
−41.099
.985
−5.857
1.00
20.87
C


ATOM
4695
C
TRP
A
314
−35.831
−2.286
−6.038
1.00
25.65
C


ATOM
4696
O
TRP
A
314
−36.069
−2.238
−7.259
1.00
25.69
O


ATOM
4698
N
ASP
A
315
−35.480
−3.400
−5.404
1.00
26.79
N


ATOM
4699
CA
ASP
A
315
−35.253
−4.667
−6.087
1.00
28.01
C


ATOM
4701
CB
ASP
A
315
−33.801
−5.097
−5.890
1.00
28.07
C


ATOM
4704
CG
ASP
A
315
−33.553
−6.536
−6.296
1.00
29.92
C


ATOM
4705
OD1
ASP
A
315
−34.262
−7.026
−7.197
1.00
32.58
O


ATOM
4706
OD2
ASP
A
315
−32.648
−7.194
−5.722
1.00
32.73
O


ATOM
4707
C
ASP
A
315
−36.202
−5.710
−5.518
1.00
28.91
C


ATOM
4708
O
ASP
A
315
−36.019
−6.163
−4.399
1.00
29.17
O


ATOM
4710
N
VAL
A
316
−37.218
−6.091
−6.283
1.00
30.25
N


ATOM
4711
CA
VAL
A
316
−38.233
−7.012
−5.778
1.00
31.47
C


ATOM
4713
CB
VAL
A
316
−39.396
−7.156
−6.755
1.00
31.56
C


ATOM
4715
CG1
VAL
A
316
−40.668
−7.590
−6.027
1.00
30.99
C


ATOM
4719
CG2
VAL
A
316
−39.033
−8.141
−7.862
1.00
31.84
C


ATOM
4723
C
VAL
A
316
−37.663
−8.405
−5.540
1.00
32.86
C


ATOM
4724
O
VAL
A
316
−38.170
−9.153
−4.708
1.00
32.99
O


ATOM
4726
N
ASN
A
317
−36.607
−8.750
−6.278
1.00
34.55
N


ATOM
4727
CA
ASN
A
317
−35.940
−10.060
−6.160
1.00
35.53
C


ATOM
4729
CB
ASN
A
317
−35.013
−10.298
−7.367
1.00
35.68
C


ATOM
4732
CG
ASN
A
317
−35.752
−10.264
−8.713
1.00
35.92
C


ATOM
4733
OD1
ASN
A
317
−36.634
−11.092
−8.971
1.00
36.95
O


ATOM
4734
ND2
ASN
A
317
−35.368
−9.323
−9.585
1.00
34.17
N


ATOM
4737
C
ASN
A
317
−35.126
−10.225
−4.871
1.00
36.40
C


ATOM
4738
O
ASN
A
317
−34.385
−11.195
−4.747
1.00
36.47
O


ATOM
4740
N
ALA
A
318
−35.239
−9.265
−3.944
1.00
37.52
N


ATOM
4741
CA
ALA
A
318
−34.614
−9.335
−2.613
1.00
38.45
C


ATOM
4743
CB
ALA
A
318
−33.371
−8.465
−2.557
1.00
38.43
C


ATOM
4747
C
ALA
A
318
−35.631
−8.902
−1.554
1.00
39.28
C


ATOM
4748
O
ALA
A
318
−35.361
−8.094
−.662
1.00
39.55
O


ATOM
4750
N
ILE
A
319
−36.823
−9.456
−1.698
1.00
40.17
N


ATOM
4751
CA
ILE
A
319
−37.905
−9.310
−.740
1.00
40.49
C


ATOM
4753
CB
ILE
A
319
−39.275
−9.588
−1.469
1.00
40.64
C


ATOM
4755
CG1
ILE
A
319
−40.473
−9.078
−.683
1.00
40.88
C


ATOM
4758
CD1
ILE
A
319
−41.799
−9.535
−1.280
1.00
40.86
C


ATOM
4762
CG2
ILE
A
319
−39.460
−11.082
−1.801
1.00
40.50
C


ATOM
4766
C
ILE
A
319
−37.656
−10.319
.393
1.00
40.69
C


ATOM
4767
O
ILE
A
319
−38.136
−10.138
1.504
1.00
40.88
O


ATOM
4769
N
ASN
A
320
−36.886
−11.374
.104
1.00
40.79
N


ATOM
4770
CA
ASN
A
320
−36.689
−12.483
1.049
1.00
40.64
C


ATOM
4772
CB
ASN
A
320
−36.240
−13.761
.314
1.00
40.73
C


ATOM
4775
CG
ASN
A
320
−37.370
−14.422
−.468
1.00
40.98
C


ATOM
4776
OD1
ASN
A
320
−38.556
−14.309
−.119
1.00
40.77
O


ATOM
4777
ND2
ASN
A
320
−37.001
−15.131
−1.529
1.00
41.31
N


ATOM
4780
C
ASN
A
320
−35.711
−12.175
2.169
1.00
40.16
C


ATOM
4781
O
ASN
A
320
−35.546
−12.978
3.077
1.00
40.15
O


ATOM
4783
N
ASP
A
321
−35.067
−11.017
2.101
1.00
39.66
N


ATOM
4784
CA
ASP
A
321
−34.111
−10.615
3.126
1.00
39.51
C


ATOM
4786
CB
ASP
A
321
−33.114
−9.571
2.575
1.00
40.17
C


ATOM
4789
CG
ASP
A
321
−32.595
−9.904
1.152
1.00
42.12
C


ATOM
4790
OD1
ASP
A
321
−32.425
−11.115
.820
1.00
44.15
O


ATOM
4791
OD2
ASP
A
321
−32.354
−8.934
.375
1.00
43.28
O


ATOM
4792
C
ASP
A
321
−34.851
−10.030
4.337
1.00
38.27
C


ATOM
4793
O
ASP
A
321
−34.304
−9.987
5.443
1.00
38.15
O


ATOM
4795
N
LEU
A
322
−36.089
−9.583
4.108
1.00
36.72
N


ATOM
4796
CA
LEU
A
322
−36.887
−8.875
5.104
1.00
35.38
C


ATOM
4798
CB
LEU
A
322
−37.894
−7.942
4.422
1.00
35.15
C


ATOM
4801
CG
LEU
A
322
−37.370
−6.840
3.503
1.00
34.83
C


ATOM
4803
CD1
LEU
A
322
−38.479
−6.286
2.626
1.00
34.45
C


ATOM
4807
CD2
LEU
A
322
−36.742
−5.740
4.316
1.00
34.89
C


ATOM
4811
C
LEU
A
322
−37.683
−9.834
5.963
1.00
34.57
C


ATOM
4812
O
LEU
A
322
−37.975
−10.933
5.527
1.00
34.27
O


ATOM
4814
N
PRO
A
323
−38.039
−9.402
7.189
1.00
33.93
N


ATOM
4815
CA
PRO
A
323
−39.070
−9.917
8.067
1.00
33.49
C


ATOM
4817
CB
PRO
A
323
−39.151
−8.840
9.141
1.00
33.31
C


ATOM
4820
CG
PRO
A
323
−37.791
−8.419
9.311
1.00
33.67
C


ATOM
4823
CD
PRO
A
323
−37.154
−8.496
7.941
1.00
34.20
C


ATOM
4826
C
PRO
A
323
−40.429
−10.036
7.425
1.00
33.33
C


ATOM
4827
O
PRO
A
323
−40.776
−9.232
6.579
1.00
33.35
O


ATOM
4828
N
ASP
A
324
−41.209
−11.005
7.891
1.00
33.37
N


ATOM
4829
CA
ASP
A
324
−42.511
−11.329
7.313
1.00
33.36
C


ATOM
4831
CB
ASP
A
324
−43.137
−12.542
8.037
1.00
33.59
C


ATOM
4834
CG
ASP
A
324
−42.496
−13.881
7.619
1.00
34.18
C


ATOM
4835
OD1
ASP
A
324
−41.885
−13.915
6.518
1.00
36.44
O


ATOM
4836
OD2
ASP
A
324
−42.607
−14.887
8.371
1.00
32.19
O


ATOM
4837
C
ASP
A
324
−43.484
−10.149
7.289
1.00
32.90
C


ATOM
4838
O
ASP
A
324
−44.108
−9.885
6.255
1.00
33.36
O


ATOM
4840
N
TYR
A
325
−43.606
−9.423
8.392
1.00
32.10
N


ATOM
4841
CA
TYR
A
325
−44.515
−8.279
8.400
1.00
31.65
C


ATOM
4843
CB
TYR
A
325
−44.718
−7.726
9.815
1.00
31.68
C


ATOM
4846
CG
TYR
A
325
−43.618
−6.846
10.352
1.00
31.35
C


ATOM
4847
CD1
TYR
A
325
−42.507
−7.389
10.992
1.00
31.31
C


ATOM
4849
CE1
TYR
A
325
−41.497
−6.574
11.504
1.00
31.46
C


ATOM
4851
CZ
TYR
A
325
−41.613
−5.192
11.392
1.00
32.41
C


ATOM
4852
OH
TYR
A
325
−40.637
−4.336
11.893
1.00
32.78
O


ATOM
4854
CE2
TYR
A
325
−42.723
−4.644
10.769
1.00
32.17
C


ATOM
4856
CD2
TYR
A
325
−43.713
−5.470
10.261
1.00
31.47
C


ATOM
4858
C
TYR
A
325
−44.094
−7.179
7.424
1.00
31.17
C


ATOM
4859
O
TYR
A
325
−44.947
−6.458
6.920
1.00
31.09
O


ATOM
4861
N
MET
A
326
−42.796
−7.068
7.145
1.00
30.73
N


ATOM
4862
CA
MET
A
326
−42.277
−6.036
6.225
1.00
30.49
C


ATOM
4864
CB
MET
A
326
−40.832
−5.702
6.565
1.00
30.16
C


ATOM
4867
CG
MET
A
326
−40.725
−4.918
7.830
1.00
29.57
C


ATOM
4870
SD
MET
A
326
−39.057
−4.376
8.166
1.00
28.39
S


ATOM
4871
CE
MET
A
326
−39.407
−2.787
8.933
1.00
26.18
C


ATOM
4875
C
MET
A
326
−42.371
−6.418
4.748
1.00
30.59
C


ATOM
4876
O
MET
A
326
−42.786
−5.603
3.920
1.00
30.48
O


ATOM
4878
N
LYS
A
327
−41.936
−7.643
4.438
1.00
30.68
N


ATOM
4879
CA
LYS
A
327
−42.170
−8.299
3.143
1.00
30.56
C


ATOM
4881
CB
LYS
A
327
−42.267
−9.830
3.326
1.00
30.84
C


ATOM
4884
CG
LYS
A
327
−41.052
−10.607
2.844
1.00
32.35
C


ATOM
4887
CD
LYS
A
327
−40.990
−12.019
3.412
1.00
34.43
C


ATOM
4890
CE
LYS
A
327
−40.221
−12.941
2.462
1.00
35.77
C


ATOM
4893
NZ
LYS
A
327
−39.691
−14.162
3.150
1.00
37.20
N


ATOM
4897
C
LYS
A
327
−43.453
−7.811
2.515
1.00
29.89
C


ATOM
4898
O
LYS
A
327
−43.447
−7.220
1.436
1.00
29.85
O


ATOM
4900
N
LEU
A
328
−44.544
−8.046
3.230
1.00
29.10
N


ATOM
4901
CA
LEU
A
328
−45.871
−7.781
2.730
1.00
28.77
C


ATOM
4903
CB
LEU
A
328
−46.899
−8.336
3.709
1.00
28.81
C


ATOM
4906
CG
LEU
A
328
−48.349
−8.364
3.257
1.00
28.82
C


ATOM
4908
CD1
LEU
A
328
−48.519
−9.260
2.046
1.00
29.09
C


ATOM
4912
CD2
LEU
A
328
−49.202
−8.845
4.411
1.00
29.16
C


ATOM
4916
C
LEU
A
328
−46.057
−6.291
2.564
1.00
28.61
C


ATOM
4917
O
LEU
A
328
−46.582
−5.828
1.554
1.00
29.00
O


ATOM
4919
N
CYS
A
329
−45.612
−5.532
3.557
1.00
28.24
N


ATOM
4920
CA
CYS
A
329
−45.737
−4.086
3.504
1.00
27.95
C


ATOM
4922
CB
CYS
A
329
−45.311
−3.459
4.834
1.00
28.21
C


ATOM
4925
SG
CYS
A
329
−45.280
−1.630
4.817
1.00
32.39
S


ATOM
4927
C
CYS
A
329
−44.921
−3.541
2.327
1.00
26.34
C


ATOM
4928
O
CYS
A
329
−45.459
−2.853
1.475
1.00
26.23
O


ATOM
4930
N
PHE
A
330
−43.642
−3.882
2.269
1.00
24.82
N


ATOM
4931
CA
PHE
A
330
−42.790
−3.514
1.130
1.00
23.82
C


ATOM
4933
CB
PHE
A
330
−41.397
−4.137
1.283
1.00
23.64
C


ATOM
4936
CG
PHE
A
330
−40.492
−3.873
.117
1.00
22.63
C


ATOM
4937
CD1
PHE
A
330
−39.845
−2.658
−.008
1.00
21.68
C


ATOM
4939
CE1
PHE
A
330
−39.020
−2.393
−1.082
1.00
21.07
C


ATOM
4941
CZ
PHE
A
330
−38.829
−3.343
−2.046
1.00
22.00
C


ATOM
4943
CE2
PHE
A
330
−39.474
−4.568
−1.944
1.00
22.60
C


ATOM
4945
CD2
PHE
A
330
−40.309
−4.824
−.865
1.00
22.36
C


ATOM
4947
C
PHE
A
330
−43.348
−3.863
−.281
1.00
22.99
C


ATOM
4948
O
PHE
A
330
−43.350
−3.012
−1.182
1.00
22.98
O


ATOM
4950
N
LEU
A
331
−43.789
−5.101
−.492
1.00
21.65
N


ATOM
4951
CA
LEU
A
331
−44.306
−5.484
−1.816
1.00
20.74
C


ATOM
4953
CB
LEU
A
331
−44.573
−6.990
−1.912
1.00
20.57
C


ATOM
4956
CG
LEU
A
331
−44.959
−7.575
−3.277
1.00
19.82
C


ATOM
4958
CD1
LEU
A
331
−43.936
−7.246
−4.329
1.00
19.00
C


ATOM
4962
CD2
LEU
A
331
−45.128
−9.092
−3.165
1.00
19.11
C


ATOM
4966
C
LEU
A
331
−45.568
−4.710
−2.159
1.00
19.98
C


ATOM
4967
O
LEU
A
331
−45.753
−4.324
−3.300
1.00
20.15
O


ATOM
4969
N
ALA
A
332
−46.431
−4.495
−1.172
1.00
19.09
N


ATOM
4970
CA
ALA
A
332
−47.619
−3.675
−1.353
1.00
18.46
C


ATOM
4972
CB
ALA
A
332
−48.406
−3.587
−.045
1.00
18.20
C


ATOM
4976
C
ALA
A
332
−47.248
−2.279
−1.856
1.00
17.91
C


ATOM
4977
O
ALA
A
332
−47.890
−1.745
−2.744
1.00
17.98
O


ATOM
4979
N
LEU
A
333
−46.197
−1.706
−1.295
1.00
17.62
N


ATOM
4980
CA
LEU
A
333
−45.753
−.353
−1.637
1.00
17.60
C


ATOM
4982
CB
LEU
A
333
−44.725
.132
−.598
1.00
17.59
C


ATOM
4985
CG
LEU
A
333
−44.122
1.533
−.761
1.00
17.16
C


ATOM
4987
CD1
LEU
A
333
−45.166
2.629
−.554
1.00
16.99
C


ATOM
4991
CD2
LEU
A
333
−42.979
1.704
.200
1.00
15.89
C


ATOM
4995
C
LEU
A
333
−45.100
−.320
−3.005
1.00
17.70
C


ATOM
4996
O
LEU
A
333
−45.321
.603
−3.795
1.00
17.86
O


ATOM
4998
N
TYR
A
334
−44.248
−1.319
−3.234
1.00
17.63
N


ATOM
4999
CA
TYR
A
334
−43.542
−1.531
−4.489
1.00
17.44
C


ATOM
5001
CB
TYR
A
334
−42.893
−2.908
−4.444
1.00
17.50
C


ATOM
5004
CG
TYR
A
334
−41.897
−3.169
−5.523
1.00
18.03
C


ATOM
5005
CD1
TYR
A
334
−40.698
−2.473
−5.569
1.00
18.83
C


ATOM
5007
CE1
TYR
A
334
−39.764
−2.725
−6.542
1.00
18.13
C


ATOM
5009
CZ
TYR
A
334
−40.016
−3.678
−7.478
1.00
18.78
C


ATOM
5010
OH
TYR
A
334
−39.096
−3.929
−8.439
1.00
21.71
O


ATOM
5012
CE2
TYR
A
334
−41.180
−4.391
−7.462
1.00
19.74
C


ATOM
5014
CD2
TYR
A
334
−42.118
−4.139
−6.472
1.00
19.59
C


ATOM
5016
C
TYR
A
334
−44.500
−1.497
−5.649
1.00
17.43
C


ATOM
5017
O
TYR
A
334
−44.264
−.803
−6.636
1.00
17.64
O


ATOM
5019
N
ASN
A
335
−45.589
−2.253
−5.504
1.00
17.30
N


ATOM
5020
CA
ASN
A
335
−46.574
−2.448
−6.553
1.00
17.23
C


ATOM
5022
CB
ASN
A
335
−47.544
−3.551
−6.166
1.00
17.32
C


ATOM
5025
CG
ASN
A
335
−46.952
−4.920
−6.332
1.00
18.16
C


ATOM
5026
OD1
ASN
A
335
−45.913
−5.090
−6.989
1.00
18.99
O


ATOM
5027
ND2
ASN
A
335
−47.616
−5.921
−5.749
1.00
18.49
N


ATOM
5030
C
ASN
A
335
−47.365
−1.218
−6.812
1.00
17.12
C


ATOM
5031
O
ASN
A
335
−47.613
−.852
−7.965
1.00
17.36
O


ATOM
5033
N
THR
A
336
−47.789
−.602
−5.722
1.00
17.20
N


ATOM
5034
CA
THR
A
336
−48.601
.593
−5.779
1.00
17.36
C


ATOM
5036
CB
THR
A
336
−48.888
1.104
−4.381
1.00
17.20
C


ATOM
5038
OG1
THR
A
336
−49.611
.103
−3.657
1.00
16.26
O


ATOM
5040
CG2
THR
A
336
−49.688
2.384
−4.452
1.00
17.18
C


ATOM
5044
C
THR
A
336
−47.893
1.691
−6.550
1.00
17.84
C


ATOM
5045
O
THR
A
336
−48.511
2.388
−7.360
1.00
18.07
O


ATOM
5047
N
ILE
A
337
−46.595
1.831
−6.298
1.00
18.14
N


ATOM
5048
CA
ILE
A
337
−45.817
2.904
−6.895
1.00
18.50
C


ATOM
5050
CB
ILE
A
337
−44.584
3.209
−6.066
1.00
18.31
C


ATOM
5052
CG1
ILE
A
337
−45.014
3.997
−4.837
1.00
18.58
C


ATOM
5055
CD1
ILE
A
337
−44.043
3.866
−3.735
1.00
20.66
C


ATOM
5059
CG2
ILE
A
337
−43.570
3.988
−6.867
1.00
16.98
C


ATOM
5063
C
ILE
A
337
−45.447
2.548
−8.314
1.00
19.29
C


ATOM
5064
O
ILE
A
337
−45.556
3.387
−9.214
1.00
19.17
O


ATOM
5066
N
ASN
A
338
−45.033
1.299
−8.513
1.00
20.16
N


ATOM
5067
CA
ASN
A
338
−44.861
.767
−9.864
1.00
20.93
C


ATOM
5069
CB
ASN
A
338
−44.409
−.695
−9.830
1.00
21.07
C


ATOM
5072
CG
ASN
A
338
−42.953
−.845
−9.439
1.00
21.61
C


ATOM
5073
OD1
ASN
A
338
−42.232
.143
−9.308
1.00
22.40
O


ATOM
5074
ND2
ASN
A
338
−42.509
−2.086
−9.260
1.00
21.92
N


ATOM
5077
C
ASN
A
338
−46.123
.914
−10.719
1.00
21.44
C


ATOM
5078
O
ASN
A
338
−46.022
1.202
−11.904
1.00
21.25
O


ATOM
5080
N
GLU
A
339
−47.303
.747
−10.128
1.00
22.21
N


ATOM
5081
CA
GLU
A
339
−48.532
1.000
−10.880
1.00
23.31
C


ATOM
5083
CB
GLU
A
339
−49.768
.464
−10.164
1.00
24.13
C


ATOM
5086
CG
GLU
A
339
−50.146
−.936
−10.660
1.00
28.69
C


ATOM
5089
CD
GLU
A
339
−50.939
−1.740
−9.638
1.00
35.06
C


ATOM
5090
OE1
GLU
A
339
−51.899
−1.136
−9.067
1.00
39.51
O


ATOM
5091
OE2
GLU
A
339
−50.596
−2.952
−9.414
1.00
36.64
O


ATOM
5092
C
GLU
A
339
−48.725
2.463
−11.269
1.00
22.71
C


ATOM
5093
O
GLU
A
339
−49.215
2.743
−12.372
1.00
22.48
O


ATOM
5095
N
ILE
A
340
−48.339
3.390
−10.390
1.00
22.19
N


ATOM
5096
CA
ILE
A
340
−48.406
4.813
−10.734
1.00
21.64
C


ATOM
5098
CB
ILE
A
340
−48.135
5.739
−9.538
1.00
21.33
C


ATOM
5100
CG1
ILE
A
340
−49.229
5.596
−8.482
1.00
21.29
C


ATOM
5103
CD1
ILE
A
340
−48.925
6.306
−7.150
1.00
20.00
C


ATOM
5107
CG2
ILE
A
340
−48.091
7.177
−9.982
1.00
20.28
C


ATOM
5111
C
ILE
A
340
−47.414
5.128
−11.861
1.00
21.78
C


ATOM
5112
O
ILE
A
340
−47.786
5.826
−12.818
1.00
22.13
O


ATOM
5114
N
ALA
A
341
−46.179
4.609
−11.771
1.00
21.34
N


ATOM
5115
CA
ALA
A
341
−45.147
4.892
−12.787
1.00
21.03
C


ATOM
5117
CB
ALA
A
341
−43.837
4.268
−12.406
1.00
20.55
C


ATOM
5121
C
ALA
A
341
−45.592
4.426
−14.183
1.00
21.32
C


ATOM
5122
O
ALA
A
341
−45.228
5.036
−15.217
1.00
21.11
O


ATOM
5124
N
TYR
A
342
−46.393
3.355
−14.196
1.00
21.45
N


ATOM
5125
CA
TYR
A
342
−47.008
2.861
−15.414
1.00
21.41
C


ATOM
5127
CB
TYR
A
342
−47.627
1.468
−15.208
1.00
21.31
C


ATOM
5130
CG
TYR
A
342
−48.336
.957
−16.450
1.00
19.60
C


ATOM
5131
CD1
TYR
A
342
−47.613
.463
−17.521
1.00
16.69
C


ATOM
5133
CE1
TYR
A
342
−48.231
.032
−18.643
1.00
15.50
C


ATOM
5135
CZ
TYR
A
342
−49.595
.085
−18.728
1.00
15.88
C


ATOM
5136
OH
TYR
A
342
−50.196
−.352
−19.877
1.00
16.46
O


ATOM
5138
CE2
TYR
A
342
−50.352
.558
−17.681
1.00
16.50
C


ATOM
5140
CD2
TYR
A
342
−49.725
.997
−16.556
1.00
18.17
C


ATOM
5142
C
TYR
A
342
−48.064
3.830
−15.899
1.00
22.08
C


ATOM
5143
O
TYR
A
342
−48.094
4.169
−17.048
1.00
22.07
O


ATOM
5145
N
ASP
A
343
−48.942
4.283
−15.032
1.00
23.41
N


ATOM
5146
CA
ASP
A
343
−49.970
5.211
−15.481
1.00
24.66
C


ATOM
5148
CB
ASP
A
343
−50.851
5.674
−14.318
1.00
25.10
C


ATOM
5151
CG
ASP
A
343
−51.720
4.552
−13.732
1.00
26.06
C


ATOM
5152
OD1
ASP
A
343
−52.107
3.613
−14.477
1.00
26.39
O


ATOM
5153
OD2
ASP
A
343
−52.032
4.642
−12.516
1.00
27.29
O


ATOM
5154
C
ASP
A
343
−49.316
6.420
−16.142
1.00
25.22
C


ATOM
5155
O
ASP
A
343
−49.755
6.874
−17.192
1.00
25.47
O


ATOM
5157
N
ASN
A
344
−48.260
6.936
−15.528
1.00
25.87
N


ATOM
5158
CA
ASN
A
344
−47.508
8.042
−16.127
1.00
26.41
C


ATOM
5160
CB
ASN
A
344
−46.498
8.605
−15.134
1.00
26.56
C


ATOM
5163
CG
ASN
A
344
−47.152
9.407
−14.073
1.00
26.95
C


ATOM
5164
OD1
ASN
A
344
−47.495
10.568
−14.296
1.00
29.13
O


ATOM
5165
ND2
ASN
A
344
−47.367
8.799
−12.916
1.00
26.36
N


ATOM
5168
C
ASN
A
344
−46.785
7.674
−17.416
1.00
26.53
C


ATOM
5169
O
ASN
A
344
−46.658
8.508
−18.304
1.00
26.74
O


ATOM
5171
N
LEU
A
345
−46.280
6.448
−17.510
1.00
26.56
N


ATOM
5172
CA
LEU
A
345
−45.634
6.019
−18.739
1.00
26.37
C


ATOM
5174
CB
LEU
A
345
−44.890
4.693
−18.550
1.00
26.32
C


ATOM
5177
CG
LEU
A
345
−43.995
4.339
−19.750
1.00
25.96
C


ATOM
5179
CD1
LEU
A
345
−42.706
5.133
−19.665
1.00
25.07
C


ATOM
5183
CD2
LEU
A
345
−43.724
2.835
−19.874
1.00
24.75
C


ATOM
5187
C
LEU
A
345
−46.679
5.908
−19.856
1.00
26.38
C


ATOM
5188
O
LEU
A
345
−46.435
6.335
−20.966
1.00
26.52
O


ATOM
5190
N
LYS
A
346
−47.839
5.340
−19.563
1.00
26.59
N


ATOM
5191
CA
LYS
A
346
−48.880
5.176
−20.572
1.00
26.89
C


ATOM
5193
CB
LYS
A
346
−50.065
4.367
−20.011
1.00
26.80
C


ATOM
5196
CG
LYS
A
346
−51.073
3.931
−21.062
1.00
26.26
C


ATOM
5199
CD
LYS
A
346
−52.210
3.080
−20.517
1.00
26.15
C


ATOM
5202
CE
LYS
A
346
−53.227
3.849
−19.689
1.00
26.40
C


ATOM
5205
NZ
LYS
A
346
−53.136
3.506
−18.223
1.00
27.67
N


ATOM
5209
C
LYS
A
346
−49.372
6.537
−21.071
1.00
27.52
C


ATOM
5210
O
LYS
A
346
−49.562
6.742
−22.272
1.00
27.57
O


ATOM
5212
N
ASP
A
347
−49.567
7.472
−20.148
1.00
28.10
N


ATOM
5213
CA
ASP
A
347
−50.309
8.689
−20.465
1.00
28.58
C


ATOM
5215
CB
ASP
A
347
−51.305
9.017
−19.329
1.00
28.85
C


ATOM
5218
CG
ASP
A
347
−52.426
7.950
−19.197
1.00
30.01
C


ATOM
5219
OD1
ASP
A
347
−52.827
7.349
−20.223
1.00
30.51
O


ATOM
5220
OD2
ASP
A
347
−52.910
7.704
−18.069
1.00
32.61
O


ATOM
5221
C
ASP
A
347
−49.407
9.871
−20.804
1.00
28.22
C


ATOM
5222
O
ASP
A
347
−49.778
10.710
−21.611
1.00
28.39
O


ATOM
5224
N
LYS
A
348
−48.228
9.930
−20.206
1.00
28.03
N


ATOM
5225
CA
LYS
A
348
−47.301
11.021
−20.467
1.00
28.01
C


ATOM
5227
CB
LYS
A
348
−46.785
11.648
−19.164
1.00
28.39
C


ATOM
5230
CG
LYS
A
348
−47.834
12.305
−18.257
1.00
30.18
C


ATOM
5233
CD
LYS
A
348
−47.143
12.898
−17.003
1.00
32.40
C


ATOM
5236
CE
LYS
A
348
−48.124
13.121
−15.874
1.00
33.59
C


ATOM
5239
NZ
LYS
A
348
−49.319
13.878
−16.341
1.00
35.48
N


ATOM
5243
C
LYS
A
348
−46.107
10.551
−21.260
1.00
27.25
C


ATOM
5244
O
LYS
A
348
−45.241
11.345
−21.566
1.00
27.60
O


ATOM
5246
N
GLY
A
349
−46.036
9.271
−21.583
1.00
26.47
N


ATOM
5247
CA
GLY
A
349
−44.863
8.742
−22.261
1.00
26.01
C


ATOM
5250
C
GLY
A
349
−43.559
9.109
−21.587
1.00
25.67
C


ATOM
5251
O
GLY
A
349
−42.613
9.472
−22.250
1.00
25.85
O


ATOM
5253
N
GLU
A
350
−43.498
9.032
−20.269
1.00
25.55
N


ATOM
5254
CA
GLU
A
350
−42.272
9.374
−19.555
1.00
25.76
C


ATOM
5256
CB
GLU
A
350
−42.332
10.814
−19.021
1.00
26.34
C


ATOM
5259
CG
GLU
A
350
−42.179
11.928
−20.106
1.00
29.04
C


ATOM
5262
CD
GLU
A
350
−40.745
12.101
−20.598
1.00
32.08
C


ATOM
5263
OE1
GLU
A
350
−39.853
12.170
−19.716
1.00
35.46
O


ATOM
5264
OE2
GLU
A
350
−40.516
12.174
−21.839
1.00
31.64
O


ATOM
5265
C
GLU
A
350
−42.084
8.414
−18.400
1.00
24.87
C


ATOM
5266
O
GLU
A
350
−43.067
8.046
−17.760
1.00
25.16
O


ATOM
5268
N
ASN
A
351
−40.833
8.009
−18.145
1.00
23.71
N


ATOM
5269
CA
ASN
A
351
−40.494
7.189
−16.980
1.00
22.88
C


ATOM
5271
CB
ASN
A
351
−39.351
6.229
−17.287
1.00
22.88
C


ATOM
5274
CG
ASN
A
351
−39.141
5.215
−16.184
1.00
22.98
C


ATOM
5275
OD1
ASN
A
351
−39.006
5.584
−15.035
1.00
22.68
O


ATOM
5276
ND2
ASN
A
351
−39.139
3.924
−16.529
1.00
24.38
N


ATOM
5279
C
ASN
A
351
−40.126
8.058
−15.786
1.00
22.40
C


ATOM
5280
O
ASN
A
351
−39.097
8.741
−15.785
1.00
22.66
O


ATOM
5282
N
ILE
A
352
−40.965
8.021
−14.760
1.00
21.55
N


ATOM
5283
CA
ILE
A
352
−40.770
8.860
−13.591
1.00
20.75
C


ATOM
5285
CB
ILE
A
352
−42.002
9.788
−13.360
1.00
20.95
C


ATOM
5287
CG1
ILE
A
352
−43.263
8.970
−13.022
1.00
20.82
C


ATOM
5290
CD1
ILE
A
352
−44.237
9.698
−12.122
1.00
20.21
C


ATOM
5294
CG2
ILE
A
352
−42.266
10.669
−14.582
1.00
19.85
C


ATOM
5298
C
ILE
A
352
−40.516
8.016
−12.339
1.00
20.37
C


ATOM
5299
O
ILE
A
352
−40.574
8.513
−11.224
1.00
20.55
O


ATOM
5301
N
LEU
A
353
−40.236
6.733
−12.519
1.00
19.98
N


ATOM
5302
CA
LEU
A
353
−40.061
5.828
−11.387
1.00
19.42
C


ATOM
5304
CB
LEU
A
353
−39.894
4.392
−11.870
1.00
19.27
C


ATOM
5307
CG
LEU
A
353
−39.989
3.318
−10.799
1.00
19.01
C


ATOM
5309
CD1
LEU
A
353
−41.164
3.574
−9.867
1.00
18.35
C


ATOM
5313
CD2
LEU
A
353
−40.085
1.938
−11.468
1.00
18.74
C


ATOM
5317
C
LEU
A
353
−38.890
6.240
−10.523
1.00
19.11
C


ATOM
5318
O
LEU
A
353
−39.022
6.319
−9.314
1.00
18.96
O


ATOM
5320
N
PRO
A
354
−37.746
6.549
−11.142
1.00
19.06
N


ATOM
5321
CA
PRO
A
354
−36.631
7.069
−10.367
1.00
19.29
C


ATOM
5323
CB
PRO
A
354
−35.749
7.717
−11.438
1.00
19.25
C


ATOM
5326
CG
PRO
A
354
−35.997
6.942
−12.638
1.00
19.08
C


ATOM
5329
CD
PRO
A
354
−37.409
6.472
−12.572
1.00
19.00
C


ATOM
5332
C
PRO
A
354
−37.024
8.122
−9.328
1.00
19.33
C


ATOM
5333
O
PRO
A
354
−36.534
8.057
−8.193
1.00
19.73
O


ATOM
5334
N
TYR
A
355
−37.891
9.063
−9.721
1.00
18.83
N


ATOM
5335
CA
TYR
A
355
−38.206
10.221
−8.898
1.00
18.79
C


ATOM
5337
CB
TYR
A
355
−38.836
11.359
−9.707
1.00
19.10
C


ATOM
5340
CG
TYR
A
355
−38.142
11.708
−11.009
1.00
20.15
C


ATOM
5341
CD1
TYR
A
355
−36.940
12.403
−11.027
1.00
20.78
C


ATOM
5343
CE1
TYR
A
355
−36.321
12.719
−12.232
1.00
22.19
C


ATOM
5345
CZ
TYR
A
355
−36.918
12.347
−13.438
1.00
22.19
C


ATOM
5346
OH
TYR
A
355
−36.341
12.660
−14.657
1.00
23.29
O


ATOM
5348
CE2
TYR
A
355
−38.113
11.677
−13.430
1.00
21.62
C


ATOM
5350
CD2
TYR
A
355
−38.720
11.371
−12.227
1.00
21.05
C


ATOM
5352
C
TYR
A
355
−39.155
9.864
−7.777
1.00
18.43
C


ATOM
5353
O
TYR
A
355
−39.081
10.460
−6.709
1.00
18.61
O


ATOM
5355
N
LEU
A
356
−40.058
8.917
−8.023
1.00
17.92
N


ATOM
5356
CA
LEU
A
356
−40.993
8.467
−6.994
1.00
17.38
C


ATOM
5358
CB
LEU
A
356
−42.136
7.658
−7.597
1.00
17.17
C


ATOM
5361
CG
LEU
A
356
−42.956
8.366
−8.682
1.00
17.56
C


ATOM
5363
CD1
LEU
A
356
−43.933
7.419
−9.371
1.00
17.47
C


ATOM
5367
CD2
LEU
A
356
−43.698
9.548
−8.112
1.00
18.08
C


ATOM
5371
C
LEU
A
356
−40.241
7.623
−5.978
1.00
17.10
C


ATOM
5372
O
LEU
A
356
−40.332
7.859
−4.783
1.00
17.62
O


ATOM
5374
N
THR
A
357
−39.464
6.656
−6.442
1.00
16.64
N


ATOM
5375
CA
THR
A
357
−38.775
5.769
−5.513
1.00
16.18
C


ATOM
5377
CB
THR
A
357
−38.104
4.554
−6.213
1.00
16.04
C


ATOM
5379
OG1
THR
A
357
−37.092
4.996
−7.123
1.00
15.93
O


ATOM
5381
CG2
THR
A
357
−39.142
3.732
−6.962
1.00
14.99
C


ATOM
5385
C
THR
A
357
−37.764
6.544
−4.686
1.00
16.16
C


ATOM
5386
O
THR
A
357
−37.599
6.278
−3.506
1.00
16.07
O


ATOM
5388
N
LYS
A
358
−37.107
7.524
−5.291
1.00
16.20
N


ATOM
5389
CA
LYS
A
358
−36.186
8.365
−4.543
1.00
16.46
C


ATOM
5391
CB
LYS
A
358
−35.518
9.386
−5.453
1.00
16.80
C


ATOM
5394
CG
LYS
A
358
−34.612
10.388
−4.741
1.00
18.30
C


ATOM
5397
CD
LYS
A
358
−33.352
9.738
−4.168
1.00
20.21
C


ATOM
5400
CE
LYS
A
358
−32.335
10.811
−3.768
1.00
21.96
C


ATOM
5403
NZ
LYS
A
358
−31.163
10.275
−3.019
1.00
22.95
N


ATOM
5407
C
LYS
A
358
−36.936
9.083
−3.440
1.00
16.26
C


ATOM
5408
O
LYS
A
358
−36.448
9.161
−2.320
1.00
16.15
O


ATOM
5410
N
ALA
A
359
−38.126
9.593
−3.763
1.00
16.11
N


ATOM
5411
CA
ALA
A
359
−38.937
10.332
−2.798
1.00
16.10
C


ATOM
5413
CB
ALA
A
359
−40.221
10.768
−3.406
1.00
15.71
C


ATOM
5417
C
ALA
A
359
−39.215
9.476
−1.588
1.00
16.52
C


ATOM
5418
O
ALA
A
359
−39.247
9.970
−.442
1.00
16.82
O


ATOM
5420
N
TRP
A
360
−39.398
8.187
−1.843
1.00
16.79
N


ATOM
5421
CA
TRP
A
360
−39.704
7.247
−.780
1.00
17.25
C


ATOM
5423
CB
TRP
A
360
−40.390
6.006
−1.352
1.00
17.41
C


ATOM
5426
CG
TRP
A
360
−41.852
6.129
−1.318
1.00
17.06
C


ATOM
5427
CD1
TRP
A
360
−42.664
6.451
−2.346
1.00
17.94
C


ATOM
5429
NE1
TRP
A
360
−43.967
6.489
−1.926
1.00
18.03
N


ATOM
5431
CE2
TRP
A
360
−44.002
6.193
−.592
1.00
17.72
C


ATOM
5432
CD2
TRP
A
360
−42.684
5.965
−.179
1.00
16.60
C


ATOM
5433
CE3
TRP
A
360
−42.441
5.654
1.155
1.00
16.65
C


ATOM
5435
CZ3
TRP
A
360
−43.508
5.573
2.022
1.00
16.91
C


ATOM
5437
CH2
TRP
A
360
−44.811
5.807
1.586
1.00
17.82
C


ATOM
5439
CZ2
TRP
A
360
−45.080
6.114
.282
1.00
18.49
C


ATOM
5441
C
TRP
A
360
−38.490
6.865
.073
1.00
17.52
C


ATOM
5442
O
TRP
A
360
−38.603
6.767
1.297
1.00
17.38
O


ATOM
5444
N
ALA
A
361
−37.344
6.651
−.568
1.00
17.82
N


ATOM
5445
CA
ALA
A
361
−36.122
6.336
.157
1.00
18.11
C


ATOM
5447
CB
ALA
A
361
−34.982
6.050
−.805
1.00
17.92
C


ATOM
5451
C
ALA
A
361
−35.781
7.507
1.063
1.00
18.52
C


ATOM
5452
O
ALA
A
361
−35.434
7.327
2.229
1.00
18.48
O


ATOM
5454
N
ASP
A
362
−35.911
8.711
.521
1.00
19.16
N


ATOM
5455
CA
ASP
A
362
−35.627
9.925
1.276
1.00
19.90
C


ATOM
5457
CB
ASP
A
362
−35.797
11.167
.387
1.00
20.39
C


ATOM
5460
CG
ASP
A
362
−34.596
11.424
−.530
1.00
21.84
C


ATOM
5461
OD1
ASP
A
362
−33.630
10.625
−.561
1.00
22.88
O


ATOM
5462
OD2
ASP
A
362
−34.630
12.453
−1.231
1.00
24.83
O


ATOM
5463
C
ASP
A
362
−36.532
10.039
2.510
1.00
19.91
C


ATOM
5464
O
ASP
A
362
−36.074
10.427
3.591
1.00
19.76
O


ATOM
5466
N
LEU
A
363
−37.813
9.707
2.346
1.00
19.95
N


ATOM
5467
CA
LEU
A
363
−38.747
9.688
3.478
1.00
19.85
C


ATOM
5469
CB
LEU
A
363
−40.175
9.415
3.006
1.00
19.70
C


ATOM
5472
CG
LEU
A
363
−41.219
9.293
4.123
1.00
18.58
C


ATOM
5474
CD1
LEU
A
363
−41.189
10.551
4.965
1.00
18.37
C


ATOM
5478
CD2
LEU
A
363
−42.597
9.073
3.551
1.00
16.43
C


ATOM
5482
C
LEU
A
363
−38.368
8.613
4.488
1.00
20.24
C


ATOM
5483
O
LEU
A
363
−38.314
8.875
5.691
1.00
20.52
O


ATOM
5485
N
CYS
A
364
−38.129
7.397
3.997
1.00
20.29
N


ATOM
5486
CA
CYS
A
364
−37.741
6.308
4.874
1.00
20.39
C


ATOM
5488
CB
CYS
A
364
−37.595
4.985
4.111
1.00
20.37
C


ATOM
5491
SG
CYS
A
364
−39.208
4.201
3.666
1.00
20.69
S


ATOM
5493
C
CYS
A
364
−36.467
6.683
5.646
1.00
20.59
C


ATOM
5494
O
CYS
A
364
−36.386
6.445
6.863
1.00
20.82
O


ATOM
5496
N
ASN
A
365
−35.495
7.314
4.980
1.00
20.35
N


ATOM
5497
CA
ASN
A
365
−34.282
7.716
5.697
1.00
20.14
C


ATOM
5499
CB
ASN
A
365
−33.188
8.203
4.754
1.00
20.18
C


ATOM
5502
CG
ASN
A
365
−32.359
7.064
4.184
1.00
20.55
C


ATOM
5503
OD1
ASN
A
365
−31.706
6.316
4.925
1.00
20.40
O


ATOM
5504
ND2
ASN
A
365
−32.365
6.938
2.854
1.00
21.25
N


ATOM
5507
C
ASN
A
365
−34.590
8.746
6.779
1.00
19.92
C


ATOM
5508
O
ASN
A
365
−33.997
8.685
7.857
1.00
19.94
O


ATOM
5510
N
ALA
A
366
−35.531
9.658
6.507
1.00
19.58
N


ATOM
5511
CA
ALA
A
366
−36.036
10.578
7.537
1.00
19.39
C


ATOM
5513
CB
ALA
A
366
−37.083
11.507
6.971
1.00
18.79
C


ATOM
5517
C
ALA
A
366
−36.597
9.784
8.730
1.00
19.75
C


ATOM
5518
O
ALA
A
366
−36.215
10.049
9.891
1.00
19.53
O


ATOM
5520
N
PHE
A
367
−37.460
8.797
8.447
1.00
19.69
N


ATOM
5521
CA
PHE
A
367
−37.985
7.925
9.502
1.00
20.11
C


ATOM
5523
CB
PHE
A
367
−38.952
6.857
8.967
1.00
20.47
C


ATOM
5526
CG
PHE
A
367
−40.293
7.370
8.494
1.00
21.35
C


ATOM
5527
CD1
PHE
A
367
−40.985
8.347
9.180
1.00
21.54
C


ATOM
5529
CE1
PHE
A
367
−42.224
8.775
8.732
1.00
21.47
C


ATOM
5531
CZ
PHE
A
367
−42.798
8.213
7.612
1.00
21.87
C


ATOM
5533
CE2
PHE
A
367
−42.135
7.230
6.924
1.00
22.60
C


ATOM
5535
CD2
PHE
A
367
−40.894
6.798
7.373
1.00
22.90
C


ATOM
5537
C
PHE
A
367
−36.871
7.179
10.252
1.00
20.12
C


ATOM
5538
O
PHE
A
367
−36.940
7.009
11.476
1.00
19.88
O


ATOM
5540
N
LEU
A
368
−35.868
6.696
9.516
1.00
20.30
N


ATOM
5541
CA
LEU
A
368
−34.775
5.931
10.133
1.00
20.19
C


ATOM
5543
CB
LEU
A
368
−33.783
5.413
9.085
1.00
20.08
C


ATOM
5546
CG
LEU
A
368
−32.743
4.363
9.514
1.00
19.30
C


ATOM
5548
CD1
LEU
A
368
−33.384
3.227
10.260
1.00
18.66
C


ATOM
5552
CD2
LEU
A
368
−31.968
3.807
8.319
1.00
18.16
C


ATOM
5556
C
LEU
A
368
−34.063
6.812
11.128
1.00
20.41
C


ATOM
5557
O
LEU
A
368
−33.842
6.407
12.257
1.00
20.01
O


ATOM
5559
N
GLN
A
369
−33.751
8.036
10.711
1.00
20.89
N


ATOM
5560
CA
GLN
A
369
−33.037
8.970
11.564
1.00
21.49
C


ATOM
5562
CB
GLN
A
369
−32.782
10.280
10.832
1.00
21.58
C


ATOM
5565
CG
GLN
A
369
−32.071
11.359
11.677
1.00
21.20
C


ATOM
5568
CD
GLN
A
369
−30.639
11.006
11.976
1.00
20.04
C


ATOM
5569
OE1
GLN
A
369
−30.282
10.650
13.108
1.00
19.10
O


ATOM
5570
NE2
GLN
A
369
−29.803
11.095
10.956
1.00
18.75
N


ATOM
5573
C
GLN
A
369
−33.763
9.265
12.870
1.00
22.21
C


ATOM
5574
O
GLN
A
369
−33.122
9.343
13.909
1.00
22.41
O


ATOM
5576
N
GLU
A
370
−35.080
9.455
12.822
1.00
23.05
N


ATOM
5577
CA
GLU
A
370
−35.856
9.718
14.046
1.00
23.71
C


ATOM
5579
CB
GLU
A
370
−37.329
10.041
13.726
1.00
24.04
C


ATOM
5582
CG
GLU
A
370
−37.484
11.293
12.862
1.00
26.84
C


ATOM
5585
CD
GLU
A
370
−38.897
11.910
12.834
1.00
30.36
C


ATOM
5586
OE1
GLU
A
370
−39.886
11.161
12.586
1.00
31.90
O


ATOM
5587
OE2
GLU
A
370
−38.992
13.164
13.014
1.00
31.62
O


ATOM
5588
C
GLU
A
370
−35.755
8.523
14.994
1.00
23.60
C


ATOM
5589
O
GLU
A
370
−35.534
8.689
16.199
1.00
23.33
O


ATOM
5591
N
ALA
A
371
−35.904
7.322
14.435
1.00
23.72
N


ATOM
5592
CA
ALA
A
371
−35.771
6.091
15.201
1.00
23.91
C


ATOM
5594
CB
ALA
A
371
−35.991
4.868
14.312
1.00
23.76
C


ATOM
5598
C
ALA
A
371
−34.392
6.051
15.840
1.00
24.20
C


ATOM
5599
O
ALA
A
371
−34.277
5.804
17.035
1.00
24.54
O


ATOM
5601
N
LYS
A
372
−33.355
6.333
15.049
1.00
24.38
N


ATOM
5602
CA
LYS
A
372
−31.979
6.273
15.530
1.00
24.39
C


ATOM
5604
CB
LYS
A
372
−30.970
6.466
14.393
1.00
24.43
C


ATOM
5607
CG
LYS
A
372
−30.623
5.164
13.645
1.00
25.20
C


ATOM
5610
CD
LYS
A
372
−29.188
5.149
13.069
1.00
26.09
C


ATOM
5613
CE
LYS
A
372
−29.114
5.437
11.557
1.00
27.01
C


ATOM
5616
NZ
LYS
A
372
−29.002
4.200
10.709
1.00
26.82
N


ATOM
5620
C
LYS
A
372
−31.717
7.268
16.645
1.00
24.59
C


ATOM
5621
O
LYS
A
372
−31.096
6.908
17.627
1.00
25.13
O


ATOM
5623
N
TRP
A
373
−32.181
8.507
16.520
1.00
24.78
N


ATOM
5624
CA
TRP
A
373
−32.006
9.473
17.610
1.00
24.90
C


ATOM
5626
CB
TRP
A
373
−32.565
10.863
17.266
1.00
24.75
C


ATOM
5629
CG
TRP
A
373
−31.701
11.677
16.338
1.00
23.59
C


ATOM
5630
CD1
TRP
A
373
−30.344
11.663
16.259
1.00
22.38
C


ATOM
5632
NE1
TRP
A
373
−29.915
12.539
15.298
1.00
21.59
N


ATOM
5634
CE2
TRP
A
373
−30.999
13.162
14.744
1.00
21.69
C


ATOM
5635
CD2
TRP
A
373
−32.147
12.644
15.376
1.00
22.41
C


ATOM
5636
CE3
TRP
A
373
−33.409
13.110
14.976
1.00
21.98
C


ATOM
5638
CZ3
TRP
A
373
−33.480
14.072
13.979
1.00
22.08
C


ATOM
5640
CH2
TRP
A
373
−32.313
14.567
13.366
1.00
22.31
C


ATOM
5642
CZ2
TRP
A
373
−31.067
14.124
13.736
1.00
21.87
C


ATOM
5644
C
TRP
A
373
−32.678
8.970
18.881
1.00
25.53
C


ATOM
5645
O
TRP
A
373
−32.101
9.072
19.972
1.00
25.99
O


ATOM
5647
N
LEU
A
374
−33.881
8.420
18.739
1.00
25.84
N


ATOM
5648
CA
LEU
A
374
−34.667
7.980
19.893
1.00
26.29
C


ATOM
5650
CB
LEU
A
374
−36.070
7.562
19.443
1.00
26.32
C


ATOM
5653
CG
LEU
A
374
−37.227
7.646
20.444
1.00
26.39
C


ATOM
5655
CD1
LEU
A
374
−38.456
8.286
19.761
1.00
26.95
C


ATOM
5659
CD2
LEU
A
374
−37.583
6.281
21.050
1.00
26.24
C


ATOM
5663
C
LEU
A
374
−33.982
6.824
20.623
1.00
26.77
C


ATOM
5664
O
LEU
A
374
−33.995
6.758
21.860
1.00
26.95
O


ATOM
5666
N
TYR
A
375
−33.383
5.919
19.854
1.00
27.23
N


ATOM
5667
CA
TYR
A
375
−32.706
4.761
20.425
1.00
27.74
C


ATOM
5669
CB
TYR
A
375
−32.195
3.814
19.328
1.00
27.78
C


ATOM
5672
CG
TYR
A
375
−31.526
2.556
19.848
1.00
28.74
C


ATOM
5673
CD1
TYR
A
375
−32.264
1.394
20.090
1.00
29.61
C


ATOM
5675
CE1
TYR
A
375
−31.652
.232
20.573
1.00
29.89
C


ATOM
5677
CZ
TYR
A
375
−30.287
.226
20.817
1.00
30.15
C


ATOM
5678
OH
TYR
A
375
−29.676
−.917
21.286
1.00
30.57
O


ATOM
5680
CE2
TYR
A
375
−29.530
1.367
20.579
1.00
30.05
C


ATOM
5682
CD2
TYR
A
375
−30.152
2.522
20.096
1.00
29.74
C


ATOM
5684
C
TYR
A
375
−31.553
5.248
21.275
1.00
27.97
C


ATOM
5685
O
TYR
A
375
−31.404
4.837
22.422
1.00
28.00
O


ATOM
5687
N
ASN
A
376
−30.763
6.158
20.719
1.00
28.32
N


ATOM
5688
CA
ASN
A
376
−29.531
6.590
21.368
1.00
28.71
C


ATOM
5690
CB
ASN
A
376
−28.569
7.161
20.329
1.00
28.60
C


ATOM
5693
CG
ASN
A
376
−28.215
6.159
19.255
1.00
28.32
C


ATOM
5694
OD1
ASN
A
376
−27.961
4.977
19.527
1.00
25.72
O


ATOM
5695
ND2
ASN
A
376
−28.195
6.631
18.015
1.00
29.29
N


ATOM
5698
C
ASN
A
376
−29.728
7.617
22.484
1.00
29.05
C


ATOM
5699
O
ASN
A
376
−28.752
8.021
23.136
1.00
29.03
O


ATOM
5701
N
LYS
A
377
−30.977
8.021
22.716
1.00
29.16
N


ATOM
5702
CA
LYS
A
377
−31.254
9.178
23.549
1.00
29.27
C


ATOM
5704
CB
LYS
A
377
−31.051
8.879
25.050
1.00
29.52
C


ATOM
5707
CG
LYS
A
377
−32.202
8.112
25.723
1.00
30.36
C


ATOM
5710
CD
LYS
A
377
−32.202
6.638
25.340
1.00
31.54
C


ATOM
5713
CE
LYS
A
377
−33.322
5.862
26.031
1.00
32.30
C


ATOM
5716
NZ
LYS
A
377
−33.559
4.509
25.411
1.00
32.13
N


ATOM
5720
C
LYS
A
377
−30.337
10.299
23.080
1.00
28.92
C


ATOM
5721
O
LYS
A
377
−29.590
10.867
23.875
1.00
28.94
O


ATOM
5723
N
SER
A
378
−30.377
10.575
21.776
1.00
28.52
N


ATOM
5724
CA
SER
A
378
−29.652
11.700
21.201
1.00
28.30
C


ATOM
5726
CB
SER
A
378
−29.623
11.620
19.678
1.00
28.33
C


ATOM
5729
OG
SER
A
378
−28.919
10.482
19.237
1.00
29.36
O


ATOM
5731
C
SER
A
378
−30.355
12.977
21.594
1.00
27.97
C


ATOM
5732
O
SER
A
378
−31.483
12.947
22.108
1.00
27.91
O


ATOM
5734
N
THR
A
379
−29.684
14.098
21.344
1.00
27.60
N


ATOM
5735
CA
THR
A
379
−30.257
15.422
21.581
1.00
27.24
C


ATOM
5737
CB
THR
A
379
−29.929
15.950
23.002
1.00
27.23
C


ATOM
5739
OG1
THR
A
379
−28.512
16.102
23.158
1.00
26.85
O


ATOM
5741
CG2
THR
A
379
−30.467
15.007
24.067
1.00
27.48
C


ATOM
5745
C
THR
A
379
−29.738
16.414
20.548
1.00
26.83
C


ATOM
5746
O
THR
A
379
−28.834
17.190
20.844
1.00
26.69
O


ATOM
5748
N
PRO
A
380
−30.305
16.390
19.331
1.00
26.56
N


ATOM
5749
CA
PRO
A
380
−29.884
17.315
18.278
1.00
26.52
C


ATOM
5751
CB
PRO
A
380
−30.380
16.648
16.992
1.00
26.46
C


ATOM
5754
CG
PRO
A
380
−31.281
15.533
17.408
1.00
26.41
C


ATOM
5757
CD
PRO
A
380
−31.395
15.508
18.887
1.00
26.53
C


ATOM
5760
C
PRO
A
380
−30.469
18.728
18.386
1.00
26.55
C


ATOM
5761
O
PRO
A
380
−31.472
18.966
19.063
1.00
26.69
O


ATOM
5762
N
THR
A
381
−29.840
19.661
17.692
1.00
26.51
N


ATOM
5763
CA
THR
A
381
−30.299
21.034
17.699
1.00
26.51
C


ATOM
5765
CB
THR
A
381
−29.261
21.955
17.033
1.00
26.99
C


ATOM
5767
OG1
THR
A
381
−28.919
21.431
15.732
1.00
27.65
O


ATOM
5769
CG2
THR
A
381
−28.003
22.082
17.930
1.00
26.41
C


ATOM
5773
C
THR
A
381
−31.635
21.167
16.970
1.00
26.06
C


ATOM
5774
O
THR
A
381
−31.972
20.347
16.112
1.00
26.03
O


ATOM
5776
N
PHE
A
382
−32.386
22.218
17.294
1.00
25.52
N


ATOM
5777
CA
PHE
A
382
−33.681
22.436
16.654
1.00
24.83
C


ATOM
5779
CB
PHE
A
382
−34.284
23.793
17.011
1.00
24.53
C


ATOM
5782
CG
PHE
A
382
−35.495
24.113
16.211
1.00
23.77
C


ATOM
5783
CD1
PHE
A
382
−36.745
23.715
16.637
1.00
24.50
C


ATOM
5785
CE1
PHE
A
382
−37.873
23.977
15.878
1.00
24.49
C


ATOM
5787
CZ
PHE
A
382
−37.744
24.630
14.673
1.00
24.41
C


ATOM
5789
CE2
PHE
A
382
−36.490
25.014
14.235
1.00
23.88
C


ATOM
5791
CD2
PHE
A
382
−35.381
24.752
14.999
1.00
23.29
C


ATOM
5793
C
PHE
A
382
−33.542
22.342
15.150
1.00
24.49
C


ATOM
5794
O
PHE
A
382
−34.361
21.731
14.485
1.00
24.32
O


ATOM
5796
N
ASP
A
383
−32.498
22.969
14.627
1.00
24.42
N


ATOM
5797
CA
ASP
A
383
−32.257
23.005
13.190
1.00
24.19
C


ATOM
5799
CB
ASP
A
383
−31.101
23.966
12.867
1.00
24.21
C


ATOM
5802
CG
ASP
A
383
−31.473
25.423
13.050
1.00
23.72
C


ATOM
5803
OD1
ASP
A
383
−32.634
25.794
12.837
1.00
25.60
O


ATOM
5804
OD2
ASP
A
383
−30.594
26.220
13.383
1.00
24.03
O


ATOM
5805
C
ASP
A
383
−31.982
21.612
12.599
1.00
24.06
C


ATOM
5806
O
ASP
A
383
−32.393
21.342
11.473
1.00
24.00
O


ATOM
5808
N
ASP
A
384
−31.298
20.739
13.336
1.00
23.78
N


ATOM
5809
CA
ASP
A
384
−31.078
19.379
12.848
1.00
24.11
C


ATOM
5811
CB
ASP
A
384
−29.981
18.651
13.632
1.00
24.54
C


ATOM
5814
CG
ASP
A
384
−28.573
19.009
13.159
1.00
26.32
C


ATOM
5815
OD1
ASP
A
384
−28.441
19.852
12.232
1.00
28.55
O


ATOM
5816
OD2
ASP
A
384
−27.599
18.451
13.728
1.00
27.18
O


ATOM
5817
C
ASP
A
384
−32.351
18.550
12.905
1.00
23.83
C


ATOM
5818
O
ASP
A
384
−32.670
17.825
11.955
1.00
24.24
O


ATOM
5820
N
TYR
A
385
−33.070
18.637
14.017
1.00
23.31
N


ATOM
5821
CA
TYR
A
385
−34.294
17.861
14.179
1.00
22.89
C


ATOM
5823
CB
TYR
A
385
−34.833
17.979
15.608
1.00
22.86
C


ATOM
5826
CG
TYR
A
385
−36.144
17.245
15.807
1.00
22.65
C


ATOM
5827
CD1
TYR
A
385
−36.163
15.868
16.030
1.00
22.36
C


ATOM
5829
CE1
TYR
A
385
−37.351
15.189
16.206
1.00
22.77
C


ATOM
5831
CZ
TYR
A
385
−38.545
15.887
16.146
1.00
23.54
C


ATOM
5832
OH
TYR
A
385
−39.733
15.212
16.309
1.00
24.38
O


ATOM
5834
CE2
TYR
A
385
−38.554
17.258
15.913
1.00
22.76
C


ATOM
5836
CD2
TYR
A
385
−37.360
17.923
15.744
1.00
22.12
C


ATOM
5838
C
TYR
A
385
−35.375
18.299
13.190
1.00
22.55
C


ATOM
5839
O
TYR
A
385
−36.050
17.467
12.584
1.00
22.85
O


ATOM
5841
N
PHE
A
386
−35.537
19.609
13.042
1.00
22.08
N


ATOM
5842
CA
PHE
A
386
−36.626
20.155
12.248
1.00
21.62
C


ATOM
5844
CB
PHE
A
386
−36.857
21.628
12.568
1.00
21.73
C


ATOM
5847
CG
PHE
A
386
−38.033
22.209
11.851
1.00
21.54
C


ATOM
5848
CD1
PHE
A
386
−39.311
21.933
12.273
1.00
21.29
C


ATOM
5850
CE1
PHE
A
386
−40.380
22.444
11.623
1.00
22.11
C


ATOM
5852
CZ
PHE
A
386
−40.193
23.240
10.517
1.00
23.28
C


ATOM
5854
CE2
PHE
A
386
−38.920
23.502
10.069
1.00
23.10
C


ATOM
5856
CD2
PHE
A
386
−37.853
22.989
10.737
1.00
22.15
C


ATOM
5858
C
PHE
A
386
−36.369
19.987
10.769
1.00
21.09
C


ATOM
5859
O
PHE
A
386
−37.278
19.700
10.006
1.00
20.91
O


ATOM
5861
N
GLY
A
387
−35.127
20.177
10.362
1.00
20.68
N


ATOM
5862
CA
GLY
A
387
−34.747
19.917
8.985
1.00
20.43
C


ATOM
5865
C
GLY
A
387
−35.151
18.521
8.544
1.00
19.95
C


ATOM
5866
O
GLY
A
387
−35.553
18.327
7.398
1.00
20.24
O


ATOM
5868
N
ASN
A
388
−35.032
17.554
9.451
1.00
19.12
N


ATOM
5869
CA
ASN
A
388
−35.451
16.182
9.202
1.00
18.73
C


ATOM
5871
CB
ASN
A
388
−34.744
15.272
10.205
1.00
18.90
C


ATOM
5874
CG
ASN
A
388
−34.863
13.795
9.871
1.00
18.35
C


ATOM
5875
OD1
ASN
A
388
−34.163
13.284
8.993
1.00
17.24
O


ATOM
5876
ND2
ASN
A
388
−35.714
13.092
10.614
1.00
17.01
N


ATOM
5879
C
ASN
A
388
−36.968
16.033
9.350
1.00
18.65
C


ATOM
5880
O
ASN
A
388
−37.624
15.370
8.551
1.00
18.82
O


ATOM
5882
N
ALA
A
389
−37.527
16.670
10.371
1.00
18.38
N


ATOM
5883
CA
ALA
A
389
−38.926
16.477
10.728
1.00
18.02
C


ATOM
5885
CB
ALA
A
389
−39.203
17.107
12.084
1.00
17.83
C


ATOM
5889
C
ALA
A
389
−39.949
16.966
9.700
1.00
17.90
C


ATOM
5890
O
ALA
A
389
−41.095
16.544
9.760
1.00
17.74
O


ATOM
5892
N
TRP
A
390
−39.585
17.860
8.783
1.00
18.11
N


ATOM
5893
CA
TRP
A
390
−40.543
18.254
7.731
1.00
18.46
C


ATOM
5895
CB
TRP
A
390
−40.459
19.737
7.329
1.00
18.59
C


ATOM
5898
CG
TRP
A
390
−39.143
20.224
6.821
1.00
19.26
C


ATOM
5899
CD1
TRP
A
390
−38.224
20.947
7.519
1.00
20.64
C


ATOM
5901
NE1
TRP
A
390
−37.138
21.224
6.729
1.00
20.62
N


ATOM
5903
CE2
TRP
A
390
−37.352
20.694
5.485
1.00
19.57
C


ATOM
5904
CD2
TRP
A
390
−38.607
20.066
5.504
1.00
19.15
C


ATOM
5905
CE3
TRP
A
390
−39.060
19.448
4.344
1.00
20.13
C


ATOM
5907
CZ3
TRP
A
390
−38.254
19.477
3.224
1.00
20.27
C


ATOM
5909
CH2
TRP
A
390
−37.017
20.110
3.239
1.00
19.38
C


ATOM
5911
CZ2
TRP
A
390
−36.549
20.724
4.356
1.00
19.49
C


ATOM
5913
C
TRP
A
390
−40.407
17.343
6.527
1.00
18.73
C


ATOM
5914
O
TRP
A
390
−41.369
17.143
5.788
1.00
18.32
O


ATOM
5916
N
LYS
A
391
−39.200
16.806
6.336
1.00
19.30
N


ATOM
5917
CA
LYS
A
391
−38.969
15.692
5.404
1.00
19.70
C


ATOM
5919
CB
LYS
A
391
−37.459
15.392
5.244
1.00
19.92
C


ATOM
5922
CG
LYS
A
391
−36.812
15.953
3.935
1.00
22.08
C


ATOM
5925
CD
LYS
A
391
−35.257
16.217
4.040
1.00
23.92
C


ATOM
5928
CE
LYS
A
391
−34.918
17.698
4.426
1.00
24.43
C


ATOM
5931
NZ
LYS
A
391
−33.699
17.901
5.307
1.00
23.23
N


ATOM
5935
C
LYS
A
391
−39.731
14.441
5.861
1.00
19.46
C


ATOM
5936
O
LYS
A
391
−40.284
13.709
5.032
1.00
20.07
O


ATOM
5938
N
SER
A
392
−39.793
14.202
7.170
1.00
18.95
N


ATOM
5939
CA
SER
A
392
−40.455
12.998
7.666
1.00
18.63
C


ATOM
5941
CB
SER
A
392
−39.850
12.508
9.000
1.00
18.75
C


ATOM
5944
OG
SER
A
392
−40.314
13.230
10.126
1.00
18.94
O


ATOM
5946
C
SER
A
392
−41.964
13.152
7.771
1.00
18.20
C


ATOM
5947
O
SER
A
392
−42.654
12.182
8.031
1.00
18.12
O


ATOM
5949
N
SER
A
393
−42.477
14.360
7.567
1.00
18.07
N


ATOM
5950
CA
SER
A
393
−43.929
14.591
7.556
1.00
18.02
C


ATOM
5952
CB
SER
A
393
−44.229
16.078
7.504
1.00
18.08
C


ATOM
5955
OG
SER
A
393
−43.995
16.558
6.192
1.00
18.01
O


ATOM
5957
C
SER
A
393
−44.594
13.971
6.340
1.00
17.87
C


ATOM
5958
O
SER
A
393
−45.778
13.661
6.372
1.00
18.03
O


ATOM
5960
N
SER
A
394
−43.823
13.841
5.264
1.00
17.69
N


ATOM
5961
CA
SER
A
394
−44.284
13.306
3.989
1.00
17.59
C


ATOM
5963
CB
SER
A
394
−45.329
12.180
4.149
1.00
17.61
C


ATOM
5966
OG
SER
A
394
−46.648
12.681
4.294
1.00
17.18
O


ATOM
5968
C
SER
A
394
−44.828
14.415
3.115
1.00
17.42
C


ATOM
5969
O
SER
A
394
−45.345
14.146
2.024
1.00
17.31
O


ATOM
5971
N
GLY
A
395
−44.711
15.654
3.592
1.00
17.23
N


ATOM
5972
CA
GLY
A
395
−45.088
16.827
2.807
1.00
17.26
C


ATOM
5975
C
GLY
A
395
−44.478
16.749
1.415
1.00
17.29
C


ATOM
5976
O
GLY
A
395
−45.203
16.653
.413
1.00
18.06
O


ATOM
5978
N
PRO
A
396
−43.145
16.754
1.331
1.00
16.72
N


ATOM
5979
CA
PRO
A
396
−42.582
16.610
.006
1.00
16.40
C


ATOM
5981
CB
PRO
A
396
−41.080
16.549
.259
1.00
16.62
C


ATOM
5984
CG
PRO
A
396
−40.903
16.749
1.775
1.00
17.11
C


ATOM
5987
CD
PRO
A
396
−42.157
17.252
2.296
1.00
16.87
C


ATOM
5990
C
PRO
A
396
−43.053
15.374
−.748
1.00
15.89
C


ATOM
5991
O
PRO
A
396
−43.501
15.498
−1.894
1.00
15.90
O


ATOM
5992
N
LEU
A
397
−42.973
14.197
−.135
1.00
15.27
N


ATOM
5993
CA
LEU
A
397
−43.287
12.976
−.886
1.00
14.57
C


ATOM
5995
CB
LEU
A
397
−43.332
11.733
−.008
1.00
14.36
C


ATOM
5998
CG
LEU
A
397
−43.541
10.431
−.781
1.00
14.02
C


ATOM
6000
CD1
LEU
A
397
−42.690
9.348
−.206
1.00
15.04
C


ATOM
6004
CD2
LEU
A
397
−44.985
9.976
−.805
1.00
13.75
C


ATOM
6008
C
LEU
A
397
−44.618
13.182
−1.542
1.00
14.39
C


ATOM
6009
O
LEU
A
397
−44.736
12.999
−2.745
1.00
14.57
O


ATOM
6011
N
GLN
A
398
−45.604
13.607
−.751
1.00
14.08
N


ATOM
6012
CA
GLN
A
398
−46.962
13.828
−1.245
1.00
13.91
C


ATOM
6014
CB
GLN
A
398
−47.860
14.363
−.136
1.00
13.99
C


ATOM
6017
CG
GLN
A
398
−48.274
13.321
.909
1.00
14.01
C


ATOM
6020
CD
GLN
A
398
−49.189
13.902
1.983
1.00
13.41
C


ATOM
6021
OE1
GLN
A
398
−49.941
14.847
1.740
1.00
14.65
O


ATOM
6022
NE2
GLN
A
398
−49.122
13.344
3.168
1.00
12.17
N


ATOM
6025
C
GLN
A
398
−47.015
14.800
−2.403
1.00
13.84
C


ATOM
6026
O
GLN
A
398
−47.677
14.547
−3.396
1.00
13.46
O


ATOM
6028
N
LEU
A
399
−46.319
15.922
−2.272
1.00
14.11
N


ATOM
6029
CA
LEU
A
399
−46.359
16.942
−3.315
1.00
14.37
C


ATOM
6031
CB
LEU
A
399
−45.900
18.282
−2.756
1.00
14.26
C


ATOM
6034
CG
LEU
A
399
−46.882
18.830
−1.704
1.00
14.44
C


ATOM
6036
CD1
LEU
A
399
−46.250
19.960
−.919
1.00
16.43
C


ATOM
6040
CD2
LEU
A
399
−48.191
19.304
−2.317
1.00
12.20
C


ATOM
6044
C
LEU
A
399
−45.582
16.525
−4.578
1.00
14.68
C


ATOM
6045
O
LEU
A
399
−46.043
16.784
−5.687
1.00
14.70
O


ATOM
6047
N
ILE
A
400
−44.443
15.844
−4.419
1.00
14.92
N


ATOM
6048
CA
ILE
A
400
−43.748
15.221
−5.564
1.00
15.08
C


ATOM
6050
CB
ILE
A
400
−42.549
14.355
−5.129
1.00
15.35
C


ATOM
6052
CG1
ILE
A
400
−41.406
15.254
−4.611
1.00
16.83
C


ATOM
6055
CD1
ILE
A
400
−40.234
14.490
−3.958
1.00
17.18
C


ATOM
6059
CG2
ILE
A
400
−42.060
13.486
−6.292
1.00
14.03
C


ATOM
6063
C
ILE
A
400
−44.682
14.329
−6.358
1.00
15.08
C


ATOM
6064
O
ILE
A
400
−44.672
14.362
−7.574
1.00
15.18
O


ATOM
6066
N
PHE
A
401
−45.474
13.524
−5.650
1.00
15.16
N


ATOM
6067
CA
PHE
A
401
−46.501
12.674
−6.258
1.00
14.90
C


ATOM
6069
CB
PHE
A
401
−47.052
11.663
−5.240
1.00
14.51
C


ATOM
6072
CG
PHE
A
401
−46.294
10.378
−5.201
1.00
12.92
C


ATOM
6073
CD1
PHE
A
401
−46.727
9.282
−5.931
1.00
11.98
C


ATOM
6075
CE1
PHE
A
401
−46.032
8.093
−5.917
1.00
11.20
C


ATOM
6077
CZ
PHE
A
401
−44.881
7.985
−5.165
1.00
12.03
C


ATOM
6079
CE2
PHE
A
401
−44.431
9.078
−4.428
1.00
11.98
C


ATOM
6081
CD2
PHE
A
401
−45.143
10.263
−4.453
1.00
12.09
C


ATOM
6083
C
PHE
A
401
−47.641
13.509
−6.806
1.00
15.39
C


ATOM
6084
O
PHE
A
401
−48.183
13.215
−7.858
1.00
15.51
O


ATOM
6086
N
ALA
A
402
−48.022
14.545
−6.080
1.00
16.15
N


ATOM
6087
CA
ALA
A
402
−49.110
15.393
−6.525
1.00
16.90
C


ATOM
6089
CB
ALA
A
402
−49.391
16.473
−5.505
1.00
16.99
C


ATOM
6093
C
ALA
A
402
−48.722
16.003
−7.856
1.00
17.50
C


ATOM
6094
O
ALA
A
402
−49.549
16.100
−8.770
1.00
17.48
O


ATOM
6096
N
TYR
A
403
−47.444
16.367
−7.963
1.00
18.28
N


ATOM
6097
CA
TYR
A
403
−46.916
17.046
−9.142
1.00
18.94
C


ATOM
6099
CB
TYR
A
403
−45.412
17.252
−9.043
1.00
19.03
C


ATOM
6102
CG
TYR
A
403
−44.823
17.801
−10.314
1.00
19.79
C


ATOM
6103
CD1
TYR
A
403
−44.973
19.138
−10.652
1.00
20.80
C


ATOM
6105
CE1
TYR
A
403
−44.436
19.643
−11.827
1.00
20.73
C


ATOM
6107
CZ
TYR
A
403
−43.759
18.804
−12.674
1.00
21.16
C


ATOM
6108
OH
TYR
A
403
−43.231
19.283
−13.836
1.00
22.98
O


ATOM
6110
CE2
TYR
A
403
−43.608
17.476
−12.370
1.00
21.23
C


ATOM
6112
CD2
TYR
A
403
−44.137
16.980
−11.195
1.00
20.93
C


ATOM
6114
C
TYR
A
403
−47.198
16.293
−10.413
1.00
19.39
C


ATOM
6115
O
TYR
A
403
−47.567
16.904
−11.422
1.00
19.57
O


ATOM
6117
N
PHE
A
404
−47.023
14.976
−10.376
1.00
19.74
N


ATOM
6118
CA
PHE
A
404
−47.239
14.177
−11.573
1.00
20.35
C


ATOM
6120
CB
PHE
A
404
−46.533
12.841
−11.466
1.00
19.96
C


ATOM
6123
CG
PHE
A
404
−45.048
12.971
−11.387
1.00
18.81
C


ATOM
6124
CD1
PHE
A
404
−44.292
13.115
−12.528
1.00
17.42
C


ATOM
6126
CE1
PHE
A
404
−42.935
13.239
−12.457
1.00
17.24
C


ATOM
6128
CZ
PHE
A
404
−42.312
13.237
−11.237
1.00
17.40
C


ATOM
6130
CE2
PHE
A
404
−43.056
13.106
−10.093
1.00
17.72
C


ATOM
6132
CD2
PHE
A
404
−44.413
12.975
−10.170
1.00
17.93
C


ATOM
6134
C
PHE
A
404
−48.713
13.994
−11.894
1.00
21.59
C


ATOM
6135
O
PHE
A
404
−49.072
13.717
−13.046
1.00
21.71
O


ATOM
6137
N
ALA
A
405
−49.572
14.169
−10.896
1.00
22.87
N


ATOM
6138
CA
ALA
A
405
−50.990
13.939
−11.099
1.00
24.06
C


ATOM
6140
CB
ALA
A
405
−51.604
13.408
−9.833
1.00
24.15
C


ATOM
6144
C
ALA
A
405
−51.724
15.192
−11.556
1.00
25.21
C


ATOM
6145
O
ALA
A
405
−52.876
15.114
−11.939
1.00
25.51
O


ATOM
6147
N
VAL
A
406
−51.056
16.336
−11.530
1.00
26.52
N


ATOM
6148
CA
VAL
A
406
−51.713
17.617
−11.759
1.00
27.68
C


ATOM
6150
CB
VAL
A
406
−51.694
18.406
−10.434
1.00
27.60
C


ATOM
6152
CG1
VAL
A
406
−51.654
19.913
−10.663
1.00
28.07
C


ATOM
6156
CG2
VAL
A
406
−52.883
18.009
−9.596
1.00
27.41
C


ATOM
6160
C
VAL
A
406
−51.097
18.429
−12.925
1.00
29.10
C


ATOM
6161
O
VAL
A
406
−51.810
19.104
−13.678
1.00
28.53
O


ATOM
6163
N
VAL
A
407
−49.772
18.356
−13.059
1.00
30.90
N


ATOM
6164
CA
VAL
A
407
−49.052
19.003
−14.151
1.00
32.15
C


ATOM
6166
CB
VAL
A
407
−47.574
19.228
−13.777
1.00
32.25
C


ATOM
6168
CG1
VAL
A
407
−46.770
19.780
−14.964
1.00
32.22
C


ATOM
6172
CG2
VAL
A
407
−47.490
20.154
−12.583
1.00
32.25
C


ATOM
6176
C
VAL
A
407
−49.134
18.160
−15.421
1.00
33.36
C


ATOM
6177
O
VAL
A
407
−48.688
17.009
−15.454
1.00
33.40
O


ATOM
6179
N
GLN
A
408
−49.693
18.767
−16.463
1.00
34.93
N


ATOM
6180
CA
GLN
A
408
−49.930
18.102
−17.749
1.00
36.16
C


ATOM
6182
CB
GLN
A
408
−50.779
19.016
−18.638
1.00
36.63
C


ATOM
6185
CG
GLN
A
408
−51.625
18.270
−19.672
1.00
38.95
C


ATOM
6188
CD
GLN
A
408
−52.991
18.930
−19.887
1.00
41.80
C


ATOM
6189
OE1
GLN
A
408
−53.739
19.150
−18.921
1.00
43.40
O


ATOM
6190
NE2
GLN
A
408
−53.323
19.244
−21.151
1.00
41.68
N


ATOM
6193
C
GLN
A
408
−48.630
17.725
−18.470
1.00
36.22
C


ATOM
6194
O
GLN
A
408
−48.456
16.588
−18.920
1.00
35.86
O


ATOM
6196
N
ASN
A
409
−47.726
18.694
−18.578
1.00
36.64
N


ATOM
6197
CA
ASN
A
409
−46.400
18.441
−19.136
1.00
36.85
C


ATOM
6199
CB
ASN
A
409
−46.134
19.306
−20.373
1.00
36.73
C


ATOM
6202
CG
ASN
A
409
−46.808
18.752
−21.604
1.00
36.19
C


ATOM
6203
OD1
ASN
A
409
−46.178
18.072
−22.420
1.00
35.00
O


ATOM
6204
ND2
ASN
A
409
−48.110
18.994
−21.720
1.00
34.99
N


ATOM
6207
C
ASN
A
409
−45.312
18.613
−18.095
1.00
36.96
C


ATOM
6208
O
ASN
A
409
−45.032
19.720
−17.618
1.00
36.81
O


ATOM
6210
N
ILE
A
410
−44.713
17.488
−17.734
1.00
37.10
N


ATOM
6211
CA
ILE
A
410
−43.625
17.508
−16.792
1.00
37.27
C


ATOM
6213
CB
ILE
A
410
−43.265
16.092
−16.295
1.00
37.38
C


ATOM
6215
CG1
ILE
A
410
−42.745
15.209
−17.426
1.00
37.84
C


ATOM
6218
CD1
ILE
A
410
−42.416
13.824
−16.985
1.00
38.96
C


ATOM
6222
CG2
ILE
A
410
−44.491
15.437
−15.676
1.00
37.67
C


ATOM
6226
C
ILE
A
410
−42.467
18.179
−17.492
1.00
37.12
C


ATOM
6227
O
ILE
A
410
−42.150
17.839
−18.617
1.00
36.87
O


ATOM
6229
N
LYS
A
411
−41.896
19.187
−16.851
1.00
37.38
N


ATOM
6230
CA
LYS
A
411
−40.680
19.813
−17.332
1.00
37.73
C


ATOM
6232
CB
LYS
A
411
−40.756
21.332
−17.171
1.00
38.15
C


ATOM
6235
CG
LYS
A
411
−41.979
21.962
−17.837
1.00
39.89
C


ATOM
6238
CD
LYS
A
411
−41.797
23.463
−18.080
1.00
42.48
C


ATOM
6241
CE
LYS
A
411
−43.029
24.069
−18.795
1.00
44.22
C


ATOM
6244
NZ
LYS
A
411
−42.977
25.576
−18.904
1.00
45.31
N


ATOM
6248
C
LYS
A
411
−39.562
19.244
−16.489
1.00
37.38
C


ATOM
6249
O
LYS
A
411
−39.721
19.112
−15.281
1.00
37.28
O


ATOM
6251
N
LYS
A
412
−38.445
18.886
−17.122
1.00
37.24
N


ATOM
6252
CA
LYS
A
412
−37.333
18.243
−16.413
1.00
36.95
C


ATOM
6254
CB
LYS
A
412
−36.286
17.701
−17.384
1.00
37.14
C


ATOM
6257
CG
LYS
A
412
−35.233
16.818
−16.720
1.00
37.89
C


ATOM
6260
CD
LYS
A
412
−34.334
16.125
−17.757
1.00
39.54
C


ATOM
6263
CE
LYS
A
412
−33.263
17.068
−18.348
1.00
40.08
C


ATOM
6266
NZ
LYS
A
412
−32.161
17.399
−17.376
1.00
40.16
N


ATOM
6270
C
LYS
A
412
−36.664
19.177
−15.419
1.00
36.42
C


ATOM
6271
O
LYS
A
412
−36.246
18.728
−14.357
1.00
36.56
O


ATOM
6273
N
GLU
A
413
−36.570
20.466
−15.744
1.00
35.77
N


ATOM
6274
CA
GLU
A
413
−35.948
21.423
−14.820
1.00
35.38
C


ATOM
6276
CB
GLU
A
413
−35.688
22.792
−15.489
1.00
35.73
C


ATOM
6279
CG
GLU
A
413
−36.510
24.013
−14.973
1.00
37.10
C


ATOM
6282
CD
GLU
A
413
−35.621
25.235
−14.633
1.00
38.51
C


ATOM
6283
OE1
GLU
A
413
−34.778
25.125
−13.714
1.00
39.24
O


ATOM
6284
OE2
GLU
A
413
−35.768
26.306
−15.266
1.00
39.04
O


ATOM
6285
C
GLU
A
413
−36.760
21.551
−13.522
1.00
34.43
C


ATOM
6286
O
GLU
A
413
−36.196
21.801
−12.460
1.00
34.34
O


ATOM
6288
N
GLU
A
414
−38.074
21.360
−13.616
1.00
33.45
N


ATOM
6289
CA
GLU
A
414
−38.956
21.407
−12.452
1.00
32.80
C


ATOM
6291
CB
GLU
A
414
−40.435
21.425
−12.864
1.00
32.83
C


ATOM
6294
CG
GLU
A
414
−40.923
22.736
−13.478
1.00
33.15
C


ATOM
6297
CD
GLU
A
414
−42.360
22.668
−14.003
1.00
33.82
C


ATOM
6298
OE1
GLU
A
414
−42.876
21.560
−14.231
1.00
35.31
O


ATOM
6299
OE2
GLU
A
414
−42.985
23.727
−14.204
1.00
33.98
O


ATOM
6300
C
GLU
A
414
−38.715
20.222
−11.536
1.00
32.18
C


ATOM
6301
O
GLU
A
414
−38.407
20.409
−10.372
1.00
32.25
O


ATOM
6303
N
ILE
A
415
−38.863
19.002
−12.044
1.00
31.63
N


ATOM
6304
CA
ILE
A
415
−38.725
17.819
−11.181
1.00
31.28
C


ATOM
6306
CB
ILE
A
415
−39.131
16.472
−11.852
1.00
31.14
C


ATOM
6308
CG1
ILE
A
415
−38.349
16.204
−13.133
1.00
31.13
C


ATOM
6311
CD1
ILE
A
415
−38.786
14.926
−13.836
1.00
30.72
C


ATOM
6315
CG2
ILE
A
415
−40.611
16.454
−12.160
1.00
31.12
C


ATOM
6319
C
ILE
A
415
−37.316
17.705
−10.636
1.00
31.08
C


ATOM
6320
O
ILE
A
415
−37.105
17.188
−9.539
1.00
31.22
O


ATOM
6322
N
GLU
A
416
−36.345
18.208
−11.381
1.00
30.67
N


ATOM
6323
CA
GLU
A
416
−34.998
18.270
−10.850
1.00
30.55
C


ATOM
6325
CB
GLU
A
416
−34.011
18.654
−11.955
1.00
30.91
C


ATOM
6328
CG
GLU
A
416
−32.654
17.967
−11.845
1.00
32.55
C


ATOM
6331
CD
GLU
A
416
−31.802
18.130
−13.109
1.00
34.91
C


ATOM
6332
OE1
GLU
A
416
−32.337
18.563
−14.161
1.00
35.53
O


ATOM
6333
OE2
GLU
A
416
−30.590
17.816
−13.052
1.00
36.45
O


ATOM
6334
C
GLU
A
416
−34.962
19.249
−9.650
1.00
29.74
C


ATOM
6335
O
GLU
A
416
−34.143
19.097
−8.738
1.00
29.73
O


ATOM
6337
N
ASN
A
417
−35.864
20.234
−9.652
1.00
28.69
N


ATOM
6338
CA
ASN
A
417
−36.060
21.134
−8.503
1.00
27.92
C


ATOM
6340
CB
ASN
A
417
−36.632
22.477
−8.963
1.00
27.75
C


ATOM
6343
CG
ASN
A
417
−35.572
23.532
−9.095
1.00
26.84
C


ATOM
6344
OD1
ASN
A
417
−35.160
24.132
−8.105
1.00
24.88
O


ATOM
6345
ND2
ASN
A
417
−35.117
23.765
−10.318
1.00
26.28
N


ATOM
6348
C
ASN
A
417
−36.917
20.580
−7.356
1.00
27.41
C


ATOM
6349
O
ASN
A
417
−36.650
20.838
−6.187
1.00
27.15
O


ATOM
6351
N
LEU
A
418
−37.955
19.834
−7.675
1.00
27.09
N


ATOM
6352
CA
LEU
A
418
−38.694
19.156
−6.628
1.00
27.02
C


ATOM
6354
CB
LEU
A
418
−39.921
18.423
−7.200
1.00
26.95
C


ATOM
6357
CG
LEU
A
418
−41.030
19.284
−7.826
1.00
25.97
C


ATOM
6359
CD1
LEU
A
418
−42.127
18.429
−8.447
1.00
24.88
C


ATOM
6363
CD2
LEU
A
418
−41.623
20.226
−6.803
1.00
24.50
C


ATOM
6367
C
LEU
A
418
−37.763
18.196
−5.860
1.00
27.26
C


ATOM
6368
O
LEU
A
418
−37.834
18.123
−4.644
1.00
27.07
O


ATOM
6370
N
GLN
A
419
−36.873
17.494
−6.561
1.00
27.67
N


ATOM
6371
CA
GLN
A
419
−35.924
16.575
−5.909
1.00
28.28
C


ATOM
6373
CB
GLN
A
419
−35.155
15.758
−6.945
1.00
28.36
C


ATOM
6376
CG
GLN
A
419
−35.747
14.375
−7.146
1.00
29.43
C


ATOM
6379
CD
GLN
A
419
−35.052
13.573
−8.215
1.00
29.76
C


ATOM
6380
OE1
GLN
A
419
−34.355
14.112
−9.066
1.00
29.93
O


ATOM
6381
NE2
GLN
A
419
−35.252
12.270
−8.184
1.00
31.33
N


ATOM
6384
C
GLN
A
419
−34.922
17.219
−4.947
1.00
28.73
C


ATOM
6385
O
GLN
A
419
−34.540
16.608
−3.952
1.00
28.90
O


ATOM
6387
N
LYS
A
420
−34.490
18.440
−5.240
1.00
29.27
N


ATOM
6388
CA
LYS
A
420
−33.608
19.166
−4.335
1.00
29.81
C


ATOM
6390
CB
LYS
A
420
−32.758
20.163
−5.129
1.00
30.24
C


ATOM
6393
CG
LYS
A
420
−31.592
19.513
−5.867
1.00
31.75
C


ATOM
6396
CD
LYS
A
420
−31.259
20.217
−7.183
1.00
34.14
C


ATOM
6399
CE
LYS
A
420
−30.140
19.463
−7.953
1.00
36.21
C


ATOM
6402
NZ
LYS
A
420
−30.073
19.759
−9.439
1.00
36.76
N


ATOM
6406
C
LYS
A
420
−34.375
19.872
−3.204
1.00
29.98
C


ATOM
6407
O
LYS
A
420
−33.759
20.547
−2.384
1.00
30.44
O


ATOM
6409
N
TYR
A
421
−35.706
19.694
−3.162
1.00
30.07
N


ATOM
6410
CA
TYR
A
421
−36.646
20.309
−2.171
1.00
29.73
C


ATOM
6412
CB
TYR
A
421
−36.298
19.949
−.712
1.00
29.59
C


ATOM
6415
CG
TYR
A
421
−36.386
18.468
−.470
1.00
30.57
C


ATOM
6416
CD1
TYR
A
421
−37.582
17.785
−.643
1.00
30.87
C


ATOM
6418
CE1
TYR
A
421
−37.665
16.414
−.448
1.00
31.28
C


ATOM
6420
CZ
TYR
A
421
−36.549
15.705
−.068
1.00
31.90
C


ATOM
6421
OH
TYR
A
421
−36.620
14.347
.124
1.00
32.43
O


ATOM
6423
CE2
TYR
A
421
−35.355
16.354
.119
1.00
32.68
C


ATOM
6425
CD2
TYR
A
421
−35.274
17.736
−.095
1.00
32.42
C


ATOM
6427
C
TYR
A
421
−36.850
21.816
−2.347
1.00
29.24
C


ATOM
6428
O
TYR
A
421
−36.709
22.590
−1.401
1.00
29.26
O


ATOM
6430
N
HIS
A
422
−37.219
22.214
−3.563
1.00
28.65
N


ATOM
6431
CA
HIS
A
422
−37.540
23.604
−3.859
1.00
28.26
C


ATOM
6433
CB
HIS
A
422
−38.150
23.715
−5.256
1.00
28.33
C


ATOM
6436
CG
HIS
A
422
−38.299
25.124
−5.748
1.00
28.44
C


ATOM
6437
ND1
HIS
A
422
−37.225
25.904
−6.106
1.00
28.97
N


ATOM
6439
CE1
HIS
A
422
−37.660
27.082
−6.520
1.00
29.40
C


ATOM
6441
NE2
HIS
A
422
−38.977
27.090
−6.443
1.00
28.14
N


ATOM
6443
CD2
HIS
A
422
−39.399
25.879
−5.965
1.00
27.76
C


ATOM
6445
C
HIS
A
422
−38.511
24.147
−2.816
1.00
27.74
C


ATOM
6446
O
HIS
A
422
−39.357
23.408
−2.310
1.00
27.53
O


ATOM
6448
N
ASP
A
423
−38.384
25.437
−2.505
1.00
27.12
N


ATOM
6449
CA
ASP
A
423
−39.186
26.072
−1.454
1.00
26.63
C


ATOM
6451
CB
ASP
A
423
−38.877
27.575
−1.349
1.00
27.05
C


ATOM
6454
CG
ASP
A
423
−37.456
27.875
−.851
1.00
28.35
C


ATOM
6455
OD1
ASP
A
423
−36.951
27.121
.018
1.00
30.99
O


ATOM
6456
OD2
ASP
A
423
−36.860
28.884
−1.322
1.00
27.89
O


ATOM
6457
C
ASP
A
423
−40.689
25.883
−1.698
1.00
25.71
C


ATOM
6458
O
ASP
A
423
−41.487
25.831
−.749
1.00
26.26
O


ATOM
6460
N
ILE
A
424
−41.075
25.781
−2.966
1.00
24.01
N


ATOM
6461
CA
ILE
A
424
−42.479
25.596
−3.327
1.00
22.72
C


ATOM
6463
CB
ILE
A
424
−42.639
25.391
−4.843
1.00
22.40
C


ATOM
6465
CG1
ILE
A
424
−44.029
25.668
−5.318
1.00
21.78
C


ATOM
6468
CD1
ILE
A
424
−44.086
25.499
−6.793
1.00
22.22
C


ATOM
6472
CG2
ILE
A
424
−42.340
23.985
−5.255
1.00
22.87
C


ATOM
6476
C
ILE
A
424
−43.103
24.442
−2.562
1.00
21.91
C


ATOM
6477
O
ILE
A
424
−44.238
24.574
−2.115
1.00
21.73
O


ATOM
6479
N
ILE
A
425
−42.365
23.334
−2.399
1.00
21.23
N


ATOM
6480
CA
ILE
A
425
−42.853
22.173
−1.634
1.00
20.73
C


ATOM
6482
CB
ILE
A
425
−42.622
20.820
−2.330
1.00
20.10
C


ATOM
6484
CG1
ILE
A
425
−41.158
20.409
−2.290
1.00
18.72
C


ATOM
6487
CD1
ILE
A
425
−40.923
19.075
−2.948
1.00
18.27
C


ATOM
6491
CG2
ILE
A
425
−43.146
20.840
−3.741
1.00
19.57
C


ATOM
6495
C
ILE
A
425
−42.252
22.060
−.242
1.00
21.21
C


ATOM
6496
O
ILE
A
425
−42.810
21.370
.613
1.00
21.30
O


ATOM
6498
N
SER
A
426
−41.129
22.721
−.002
1.00
21.56
N


ATOM
6499
CA
SER
A
426
−40.492
22.620
1.299
1.00
22.23
C


ATOM
6501
CB
SER
A
426
−39.043
23.082
1.234
1.00
22.36
C


ATOM
6504
OG
SER
A
426
−38.977
24.501
1.342
1.00
24.09
O


ATOM
6506
C
SER
A
426
−41.242
23.460
2.334
1.00
22.39
C


ATOM
6507
O
SER
A
426
−41.390
23.051
3.491
1.00
22.95
O


ATOM
6509
N
ARG
A
427
−41.701
24.643
1.937
1.00
22.08
N


ATOM
6510
CA
ARG
A
427
−42.332
25.541
2.905
1.00
21.87
C


ATOM
6512
CB
ARG
A
427
−42.466
26.955
2.345
1.00
21.99
C


ATOM
6515
CG
ARG
A
427
−41.170
27.696
2.493
1.00
23.36
C


ATOM
6518
CD
ARG
A
427
−40.954
28.727
1.433
1.00
26.07
C


ATOM
6521
NE
ARG
A
427
−39.680
29.407
1.667
1.00
28.18
N


ATOM
6523
CZ
ARG
A
427
−39.127
30.298
.844
1.00
29.21
C


ATOM
6524
NH1
ARG
A
427
−39.723
30.633
−.298
1.00
28.60
N


ATOM
6527
NH2
ARG
A
427
−37.962
30.854
1.173
1.00
30.48
N


ATOM
6530
C
ARG
A
427
−43.651
25.003
3.434
1.00
21.21
C


ATOM
6531
O
ARG
A
427
−43.817
24.891
4.645
1.00
21.11
O


ATOM
6533
N
PRO
A
428
−44.573
24.622
2.541
1.00
20.52
N


ATOM
6534
CA
PRO
A
428
−45.789
23.999
3.040
1.00
20.27
C


ATOM
6536
CB
PRO
A
428
−46.410
23.369
1.791
1.00
20.44
C


ATOM
6539
CG
PRO
A
428
−45.864
24.119
.666
1.00
20.63
C


ATOM
6542
CD
PRO
A
428
−44.521
24.644
1.074
1.00
20.54
C


ATOM
6545
C
PRO
A
428
−45.450
22.917
4.050
1.00
19.75
C


ATOM
6546
O
PRO
A
428
−46.089
22.834
5.095
1.00
19.77
O


ATOM
6547
N
SER
A
429
−44.424
22.123
3.747
1.00
18.97
N


ATOM
6548
CA
SER
A
429
−44.008
21.045
4.636
1.00
18.45
C


ATOM
6550
CB
SER
A
429
−42.954
20.180
3.972
1.00
18.35
C


ATOM
6553
OG
SER
A
429
−43.452
19.716
2.733
1.00
19.12
O


ATOM
6555
C
SER
A
429
−43.519
21.525
5.985
1.00
17.97
C


ATOM
6556
O
SER
A
429
−43.733
20.846
6.968
1.00
17.88
O


ATOM
6558
N
HIS
A
430
−42.879
22.689
6.053
1.00
17.73
N


ATOM
6559
CA
HIS
A
430
−42.593
23.289
7.359
1.00
17.71
C


ATOM
6561
CB
HIS
A
430
−41.937
24.668
7.241
1.00
17.97
C


ATOM
6564
CG
HIS
A
430
−40.558
24.663
6.651
1.00
18.87
C


ATOM
6565
ND1
HIS
A
430
−39.940
23.524
6.186
1.00
19.85
N


ATOM
6567
CE1
HIS
A
430
−38.746
23.836
5.709
1.00
19.43
C


ATOM
6569
NE2
HIS
A
430
−38.573
25.137
5.838
1.00
19.06
N


ATOM
6571
CD2
HIS
A
430
−39.693
25.681
6.418
1.00
19.32
C


ATOM
6573
C
HIS
A
430
−43.914
23.444
8.131
1.00
17.31
C


ATOM
6574
O
HIS
A
430
−44.023
23.013
9.287
1.00
17.06
O


ATOM
6576
N
ILE
A
431
−44.913
24.050
7.475
1.00
16.74
N


ATOM
6577
CA
ILE
A
431
−46.218
24.304
8.091
1.00
16.15
C


ATOM
6579
CB
ILE
A
431
−47.174
25.057
7.174
1.00
16.18
C


ATOM
6581
CG1
ILE
A
431
−46.613
26.428
6.801
1.00
17.05
C


ATOM
6584
CD1
ILE
A
431
−46.441
27.352
7.994
1.00
18.34
C


ATOM
6588
CG2
ILE
A
431
−48.496
25.263
7.860
1.00
15.28
C


ATOM
6592
C
ILE
A
431
−46.899
23.021
8.476
1.00
15.75
C


ATOM
6593
O
ILE
A
431
−47.624
22.980
9.445
1.00
15.95
O


ATOM
6595
N
PHE
A
432
−46.663
21.969
7.714
1.00
15.59
N


ATOM
6596
CA
PHE
A
432
−47.192
20.652
8.041
1.00
15.52
C


ATOM
6598
CB
PHE
A
432
−46.837
19.683
6.913
1.00
15.23
C


ATOM
6601
CG
PHE
A
432
−47.451
18.318
7.031
1.00
15.98
C


ATOM
6602
CD1
PHE
A
432
−48.324
17.972
8.052
1.00
16.55
C


ATOM
6604
CE1
PHE
A
432
−48.867
16.700
8.102
1.00
16.04
C


ATOM
6606
CZ
PHE
A
432
−48.559
15.774
7.130
1.00
15.29
C


ATOM
6608
CE2
PHE
A
432
−47.718
16.107
6.114
1.00
15.37
C


ATOM
6610
CD2
PHE
A
432
−47.172
17.364
6.061
1.00
16.74
C


ATOM
6612
C
PHE
A
432
−46.640
20.191
9.392
1.00
15.54
C


ATOM
6613
O
PHE
A
432
−47.383
20.006
10.343
1.00
15.37
O


ATOM
6615
N
ARG
A
433
−45.328
20.041
9.483
1.00
15.98
N


ATOM
6616
CA
ARG
A
433
−44.695
19.542
10.709
1.00
16.20
C


ATOM
6618
CB
ARG
A
433
−43.176
19.476
10.526
1.00
15.85
C


ATOM
6621
CG
ARG
A
433
−42.411
19.183
11.804
1.00
16.11
C


ATOM
6624
CD
ARG
A
433
−42.848
17.890
12.489
1.00
15.70
C


ATOM
6627
NE
ARG
A
433
−42.811
16.747
11.588
1.00
15.64
N


ATOM
6629
CZ
ARG
A
433
−43.377
15.571
11.838
1.00
16.27
C


ATOM
6630
NH1
ARG
A
433
−44.029
15.359
12.965
1.00
16.82
N


ATOM
6633
NH2
ARG
A
433
−43.302
14.598
10.948
1.00
16.96
N


ATOM
6636
C
ARG
A
433
−45.029
20.411
11.935
1.00
16.46
C


ATOM
6637
O
ARG
A
433
−45.296
19.890
13.031
1.00
16.31
O


ATOM
6639
N
LEU
A
434
−45.006
21.731
11.724
1.00
16.47
N


ATOM
6640
CA
LEU
A
434
−45.123
22.701
12.800
1.00
15.98
C


ATOM
6642
CB
LEU
A
434
−44.771
24.114
12.303
1.00
15.83
C


ATOM
6645
CG
LEU
A
434
−43.287
24.486
12.218
1.00
14.95
C


ATOM
6647
CD1
LEU
A
434
−43.130
25.918
11.826
1.00
15.09
C


ATOM
6651
CD2
LEU
A
434
−42.600
24.281
13.536
1.00
14.37
C


ATOM
6655
C
LEU
A
434
−46.517
22.660
13.387
1.00
16.19
C


ATOM
6656
O
LEU
A
434
−46.669
22.564
14.604
1.00
15.92
O


ATOM
6658
N
CYS
A
435
−47.529
22.715
12.520
1.00
16.78
N


ATOM
6659
CA
CYS
A
435
−48.936
22.591
12.940
1.00
17.48
C


ATOM
6661
CB
CYS
A
435
−49.870
22.617
11.726
1.00
17.54
C


ATOM
6664
SG
CYS
A
435
−50.110
24.233
10.985
1.00
18.80
S


ATOM
6666
C
CYS
A
435
−49.192
21.296
13.708
1.00
17.84
C


ATOM
6667
O
CYS
A
435
−50.043
21.250
14.597
1.00
17.92
O


ATOM
6669
N
ASN
A
436
−48.446
20.257
13.333
1.00
18.24
N


ATOM
6670
CA
ASN
A
436
−48.603
18.913
13.846
1.00
18.50
C


ATOM
6672
CB
ASN
A
436
−47.936
17.949
12.867
1.00
18.63
C


ATOM
6675
CG
ASN
A
436
−48.156
16.487
13.213
1.00
18.58
C


ATOM
6676
OD1
ASN
A
436
−48.810
16.129
14.185
1.00
18.80
O


ATOM
6677
ND2
ASN
A
436
−47.588
15.633
12.398
1.00
19.29
N


ATOM
6680
C
ASN
A
436
−47.975
18.755
15.210
1.00
18.83
C


ATOM
6681
O
ASN
A
436
−48.551
18.153
16.115
1.00
19.00
O


ATOM
6683
N
ASP
A
437
−46.763
19.255
15.351
1.00
19.29
N


ATOM
6684
CA
ASP
A
437
−46.097
19.179
16.634
1.00
19.62
C


ATOM
6686
CB
ASP
A
437
−44.615
19.522
16.510
1.00
19.45
C


ATOM
6689
CG
ASP
A
437
−43.849
18.480
15.734
1.00
19.36
C


ATOM
6690
OD1
ASP
A
437
−44.491
17.586
15.158
1.00
19.89
O


ATOM
6691
OD2
ASP
A
437
−42.608
18.543
15.699
1.00
19.86
O


ATOM
6692
C
ASP
A
437
−46.799
20.106
17.597
1.00
20.01
C


ATOM
6693
O
ASP
A
437
−46.916
19.790
18.779
1.00
20.30
O


ATOM
6695
N
LEU
A
438
−47.288
21.233
17.092
1.00
20.40
N


ATOM
6696
CA
LEU
A
438
−47.996
22.174
17.940
1.00
21.03
C


ATOM
6698
CB
LEU
A
438
−48.510
23.363
17.126
1.00
21.01
C


ATOM
6701
CG
LEU
A
438
−47.575
24.565
17.078
1.00
20.32
C


ATOM
6703
CD1
LEU
A
438
−47.938
25.524
15.952
1.00
19.14
C


ATOM
6707
CD2
LEU
A
438
−47.611
25.263
18.415
1.00
19.73
C


ATOM
6711
C
LEU
A
438
−49.153
21.496
18.679
1.00
21.81
C


ATOM
6712
O
LEU
A
438
−49.277
21.633
19.902
1.00
21.55
O


ATOM
6714
N
ALA
A
439
−49.973
20.754
17.932
1.00
22.76
N


ATOM
6715
CA
ALA
A
439
−51.138
20.055
18.486
1.00
23.58
C


ATOM
6717
CB
ALA
A
439
−51.905
19.360
17.386
1.00
23.57
C


ATOM
6721
C
ALA
A
439
−50.746
19.042
19.542
1.00
24.47
C


ATOM
6722
O
ALA
A
439
−51.386
18.951
20.575
1.00
24.63
O


ATOM
6724
N
SER
A
440
−49.685
18.291
19.276
1.00
25.78
N


ATOM
6725
CA
SER
A
440
−49.242
17.227
20.168
1.00
26.84
C


ATOM
6727
CB
SER
A
440
−48.566
16.130
19.353
1.00
26.92
C


ATOM
6730
OG
SER
A
440
−47.321
16.582
18.859
1.00
27.46
O


ATOM
6732
C
SER
A
440
−48.270
17.691
21.244
1.00
27.68
C


ATOM
6733
O
SER
A
440
−47.934
16.923
22.132
1.00
28.00
O


ATOM
6735
N
ALA
A
441
−47.822
18.935
21.174
1.00
28.83
N


ATOM
6736
CA
ALA
A
441
−46.760
19.419
22.056
1.00
29.73
C


ATOM
6738
CB
ALA
A
441
−46.529
20.910
21.841
1.00
29.84
C


ATOM
6742
C
ALA
A
441
−46.982
19.132
23.539
1.00
30.57
C


ATOM
6743
O
ALA
A
441
−46.306
18.274
24.093
1.00
30.50
O


ATOM
6745
N
SER
A
442
−47.929
19.827
24.172
1.00
31.90
N


ATOM
6746
CA
SER
A
442
−47.999
19.862
25.654
1.00
32.95
C


ATOM
6748
CB
SER
A
442
−49.043
20.870
26.169
1.00
32.91
C


ATOM
6751
OG
SER
A
442
−50.331
20.614
25.650
1.00
33.50
O


ATOM
6753
C
SER
A
442
−48.219
18.497
26.289
1.00
33.69
C


ATOM
6754
O
SER
A
442
−47.754
18.246
27.397
1.00
33.59
O


ATOM
6756
N
ALA
A
443
−48.922
17.623
25.578
1.00
34.93
N


ATOM
6757
CA
ALA
A
443
−49.050
16.231
25.980
1.00
35.81
C


ATOM
6759
CB
ALA
A
443
−49.981
15.488
25.030
1.00
35.72
C


ATOM
6763
C
ALA
A
443
−47.663
15.580
25.996
1.00
36.71
C


ATOM
6764
O
ALA
A
443
−47.198
15.115
27.047
1.00
36.94
O


ATOM
6766
N
GLU
A
444
−46.999
15.567
24.838
1.00
37.47
N


ATOM
6767
CA
GLU
A
444
−45.688
14.930
24.718
1.00
37.93
C


ATOM
6769
CB
GLU
A
444
−45.164
14.996
23.277
1.00
37.99
C


ATOM
6772
CG
GLU
A
444
−45.952
14.100
22.326
1.00
39.24
C


ATOM
6775
CD
GLU
A
444
−45.419
14.080
20.886
1.00
41.03
C


ATOM
6776
OE1
GLU
A
444
−44.612
14.958
20.510
1.00
42.16
O


ATOM
6777
OE2
GLU
A
444
−45.827
13.178
20.119
1.00
42.36
O


ATOM
6778
C
GLU
A
444
−44.694
15.534
25.708
1.00
38.09
C


ATOM
6779
O
GLU
A
444
−43.924
14.803
26.324
1.00
38.17
O


ATOM
6781
N
ILE
A
445
−44.739
16.851
25.889
1.00
38.41
N


ATOM
6782
CA
ILE
A
445
−43.829
17.530
26.814
1.00
38.75
C


ATOM
6784
CB
ILE
A
445
−43.802
19.065
26.587
1.00
38.69
C


ATOM
6786
CG1
ILE
A
445
−43.314
19.396
25.170
1.00
38.18
C


ATOM
6789
CD1
ILE
A
445
−43.702
20.775
24.712
1.00
37.58
C


ATOM
6793
CG2
ILE
A
445
−42.909
19.753
27.617
1.00
38.26
C


ATOM
6797
C
ILE
A
445
−44.243
17.194
28.246
1.00
39.35
C


ATOM
6798
O
ILE
A
445
−45.039
17.901
28.870
1.00
39.44
O


ATOM
6800
N
ALA
A
446
−43.696
16.092
28.750
1.00
40.02
N


ATOM
6801
CA
ALA
A
446
−44.082
15.540
30.045
1.00
40.51
C


ATOM
6803
CB
ALA
A
446
−43.815
16.552
31.170
1.00
40.55
C


ATOM
6807
C
ALA
A
446
−45.559
15.130
30.029
1.00
40.86
C


ATOM
6808
O
ALA
A
446
−46.407
16.021
30.121
1.00
40.77
O


ATOM
6810
N
ARG
A
447
−45.925
13.842
29.899
1.00
41.31
N


ATOM
6811
CA
ARG
A
447
−45.072
12.633
29.691
1.00
41.58
C


ATOM
6813
CB
ARG
A
447
−45.272
12.092
28.258
1.00
41.84
C


ATOM
6816
CG
ARG
A
447
−46.667
11.495
27.991
1.00
42.69
C


ATOM
6819
CD
ARG
A
447
−46.778
10.901
26.582
1.00
43.92
C


ATOM
6822
NE
ARG
A
447
−47.764
11.589
25.744
1.00
45.34
N


ATOM
6824
CZ
ARG
A
447
−47.887
11.435
24.420
1.00
46.56
C


ATOM
6825
NH1
ARG
A
447
−47.077
10.623
23.742
1.00
46.71
N


ATOM
6828
NH2
ARG
A
447
−48.828
12.107
23.757
1.00
46.97
N


ATOM
6831
C
ARG
A
447
−43.581
12.728
30.054
1.00
41.38
C


ATOM
6832
O
ARG
A
447
−43.231
12.747
31.238
1.00
41.92
O


ATOM
6834
N
GLY
A
448
−42.710
12.736
29.051
1.00
40.88
N


ATOM
6835
CA
GLY
A
448
−41.312
13.128
29.234
1.00
40.44
C


ATOM
6838
C
GLY
A
448
−40.579
13.316
27.915
1.00
40.08
C


ATOM
6839
O
GLY
A
448
−39.364
13.488
27.901
1.00
40.05
O


ATOM
6841
N
GLU
A
449
−41.323
13.332
26.809
1.00
39.56
N


ATOM
6842
CA
GLU
A
449
−40.750
13.132
25.486
1.00
39.19
C


ATOM
6844
CB
GLU
A
449
−41.827
12.668
24.502
1.00
39.52
C


ATOM
6847
CG
GLU
A
449
−42.403
11.290
24.810
1.00
40.61
C


ATOM
6850
CD
GLU
A
449
−43.348
10.782
23.715
1.00
42.21
C


ATOM
6851
OE1
GLU
A
449
−43.068
11.019
22.503
1.00
41.83
O


ATOM
6852
OE2
GLU
A
449
−44.368
10.141
24.082
1.00
42.41
O


ATOM
6853
C
GLU
A
449
−40.037
14.365
24.921
1.00
38.34
C


ATOM
6854
O
GLU
A
449
−40.511
15.497
25.031
1.00
38.01
O


ATOM
6856
N
THR
A
450
−38.904
14.106
24.279
1.00
37.37
N


ATOM
6857
CA
THR
A
450
−38.047
15.138
23.726
1.00
36.37
C


ATOM
6859
CB
THR
A
450
−36.607
14.985
24.285
1.00
36.39
C


ATOM
6861
OG1
THR
A
450
−35.846
16.153
23.982
1.00
36.63
O


ATOM
6863
CG2
THR
A
450
−35.888
13.749
23.706
1.00
36.52
C


ATOM
6867
C
THR
A
450
−38.013
15.100
22.190
1.00
35.36
C


ATOM
6868
O
THR
A
450
−37.175
15.764
21.583
1.00
35.43
O


ATOM
6870
N
ALA
A
451
−38.917
14.338
21.563
1.00
33.98
N


ATOM
6871
CA
ALA
A
451
−38.920
14.182
20.096
1.00
32.81
C


ATOM
6873
CB
ALA
A
451
−39.030
12.708
19.708
1.00
32.99
C


ATOM
6877
C
ALA
A
451
−40.043
14.987
19.459
1.00
31.46
C


ATOM
6878
O
ALA
A
451
−41.050
14.432
18.996
1.00
31.06
O


ATOM
6880
N
ASN
A
452
−39.848
16.302
19.432
1.00
29.88
N


ATOM
6881
CA
ASN
A
452
−40.881
17.225
18.973
1.00
28.81
C


ATOM
6883
CB
ASN
A
452
−41.957
17.329
20.051
1.00
28.57
C


ATOM
6886
CG
ASN
A
452
−42.969
18.380
19.757
1.00
28.30
C


ATOM
6887
OD1
ASN
A
452
−42.622
19.517
19.468
1.00
29.35
O


ATOM
6888
ND2
ASN
A
452
−44.236
18.018
19.836
1.00
28.36
N


ATOM
6891
C
ASN
A
452
−40.285
18.590
18.641
1.00
27.76
C


ATOM
6892
O
ASN
A
452
−39.365
19.018
19.296
1.00
28.04
O


ATOM
6894
N
SER
A
453
−40.807
19.269
17.630
1.00
26.77
N


ATOM
6895
CA
SER
A
453
−40.226
20.533
17.182
1.00
26.38
C


ATOM
6897
CB
SER
A
453
−40.912
21.015
15.903
1.00
26.40
C


ATOM
6900
OG
SER
A
453
−40.796
20.044
14.876
1.00
25.79
O


ATOM
6902
C
SER
A
453
−40.253
21.643
18.233
1.00
26.12
C


ATOM
6903
O
SER
A
453
−39.280
22.361
18.385
1.00
25.97
O


ATOM
6905
N
VAL
A
454
−41.368
21.784
18.942
1.00
26.15
N


ATOM
6906
CA
VAL
A
454
−41.518
22.780
20.022
1.00
26.21
C


ATOM
6908
CB
VAL
A
454
−42.975
22.818
20.532
1.00
26.04
C


ATOM
6910
CG1
VAL
A
454
−43.122
23.787
21.694
1.00
25.24
C


ATOM
6914
CG2
VAL
A
454
−43.913
23.190
19.401
1.00
26.32
C


ATOM
6918
C
VAL
A
454
−40.617
22.451
21.211
1.00
26.63
C


ATOM
6919
O
VAL
A
454
−40.173
23.328
21.959
1.00
26.53
O


ATOM
6921
N
SER
A
455
−40.374
21.160
21.380
1.00
27.21
N


ATOM
6922
CA
SER
A
455
−39.541
20.651
22.437
1.00
27.60
C


ATOM
6924
CB
SER
A
455
−39.677
19.141
22.490
1.00
27.31
C


ATOM
6927
OG
SER
A
455
−38.922
18.625
23.545
1.00
28.13
O


ATOM
6929
C
SER
A
455
−38.096
21.045
22.181
1.00
28.29
C


ATOM
6930
O
SER
A
455
−37.445
21.628
23.043
1.00
28.63
O


ATOM
6932
N
CYS
A
456
−37.599
20.748
20.988
1.00
29.04
N


ATOM
6933
CA
CYS
A
456
−36.219
21.082
20.640
1.00
29.65
C


ATOM
6935
CB
CYS
A
456
−35.868
20.535
19.256
1.00
29.52
C


ATOM
6938
SG
CYS
A
456
−35.959
18.734
19.163
1.00
29.71
S


ATOM
6940
C
CYS
A
456
−35.955
22.589
20.710
1.00
30.26
C


ATOM
6941
O
CYS
A
456
−34.887
23.007
21.162
1.00
30.49
O


ATOM
6943
N
TYR
A
457
−36.924
23.399
20.283
1.00
30.88
N


ATOM
6944
CA
TYR
A
457
−36.767
24.850
20.308
1.00
31.45
C


ATOM
6946
CB
TYR
A
457
−37.967
25.557
19.648
1.00
31.55
C


ATOM
6949
CG
TYR
A
457
−37.691
26.991
19.178
1.00
32.09
C


ATOM
6950
CD1
TYR
A
457
−37.378
27.269
17.843
1.00
31.69
C


ATOM
6952
CE1
TYR
A
457
−37.128
28.569
17.418
1.00
31.73
C


ATOM
6954
CZ
TYR
A
457
−37.186
29.613
18.328
1.00
32.84
C


ATOM
6955
OH
TYR
A
457
−36.945
30.917
17.927
1.00
33.81
O


ATOM
6957
CE2
TYR
A
457
−37.492
29.364
19.658
1.00
32.72
C


ATOM
6959
CD2
TYR
A
457
−37.746
28.064
20.074
1.00
32.44
C


ATOM
6961
C
TYR
A
457
−36.571
25.295
21.758
1.00
31.99
C


ATOM
6962
O
TYR
A
457
−35.648
26.053
22.052
1.00
31.98
O


ATOM
6964
N
MET
A
458
−37.419
24.799
22.661
1.00
32.76
N


ATOM
6965
CA
MET
A
458
−37.211
24.987
24.105
1.00
33.37
C


ATOM
6967
CB
MET
A
458
−38.151
24.097
24.921
1.00
33.43
C


ATOM
6970
CG
MET
A
458
−39.570
24.593
25.059
1.00
33.70
C


ATOM
6973
SD
MET
A
458
−40.641
23.312
25.737
1.00
34.59
S


ATOM
6974
CE
MET
A
458
−39.709
22.763
27.179
1.00
34.71
C


ATOM
6978
C
MET
A
458
−35.784
24.641
24.517
1.00
33.78
C


ATOM
6979
O
MET
A
458
−35.101
25.437
25.151
1.00
33.73
O


ATOM
6981
N
ARG
A
459
−35.347
23.445
24.145
1.00
34.46
N


ATOM
6982
CA
ARG
A
459
−34.051
22.926
24.581
1.00
35.15
C


ATOM
6984
CB
ARG
A
459
−33.876
21.447
24.171
1.00
35.68
C


ATOM
6987
CG
ARG
A
459
−32.494
20.864
24.500
1.00
37.23
C


ATOM
6990
CD
ARG
A
459
−32.491
19.345
24.752
1.00
39.80
C


ATOM
6993
NE
ARG
A
459
−33.140
18.538
23.711
1.00
42.88
N


ATOM
6995
CZ
ARG
A
459
−32.704
18.407
22.453
1.00
45.43
C


ATOM
6996
NH1
ARG
A
459
−31.615
19.062
22.035
1.00
47.50
N


ATOM
6999
NH2
ARG
A
459
−33.366
17.629
21.595
1.00
44.77
N


ATOM
7002
C
ARG
A
459
−32.878
23.749
24.072
1.00
34.92
C


ATOM
7003
O
ARG
A
459
−32.007
24.109
24.849
1.00
35.06
O


ATOM
7005
N
THR
A
460
−32.854
24.047
22.778
1.00
34.96
N


ATOM
7006
CA
THR
A
460
−31.674
24.678
22.163
1.00
34.91
C


ATOM
7008
CB
THR
A
460
−31.494
24.263
20.680
1.00
34.88
C


ATOM
7010
OG1
THR
A
460
−32.330
25.067
19.841
1.00
34.39
O


ATOM
7012
CG2
THR
A
460
−31.825
22.776
20.488
1.00
35.44
C


ATOM
7016
C
THR
A
460
−31.672
26.210
22.258
1.00
34.78
C


ATOM
7017
O
THR
A
460
−30.673
26.837
21.911
1.00
34.92
O


ATOM
7019
N
LYS
A
461
−32.781
26.806
22.701
1.00
34.41
N


ATOM
7020
CA
LYS
A
461
−32.811
28.231
23.039
1.00
34.25
C


ATOM
7022
CB
LYS
A
461
−33.921
28.947
22.265
1.00
34.56
C


ATOM
7025
CG
LYS
A
461
−33.717
28.986
20.750
1.00
35.84
C


ATOM
7028
CD
LYS
A
461
−32.519
29.859
20.347
1.00
37.64
C


ATOM
7031
CE
LYS
A
461
−32.232
29.804
18.838
1.00
38.44
C


ATOM
7034
NZ
LYS
A
461
−33.141
30.671
18.027
1.00
38.31
N


ATOM
7038
C
LYS
A
461
−32.974
28.476
24.544
1.00
33.73
C


ATOM
7039
O
LYS
A
461
−32.994
29.626
24.983
1.00
33.23
O


ATOM
7041
N
GLY
A
462
−33.078
27.395
25.321
1.00
33.48
N


ATOM
7042
CA
GLY
A
462
−33.173
27.464
26.784
1.00
33.23
C


ATOM
7045
C
GLY
A
462
−34.332
28.320
27.239
1.00
32.95
C


ATOM
7046
O
GLY
A
462
−34.144
29.258
28.004
1.00
33.17
O


ATOM
7048
N
ILE
A
463
−35.528
28.000
26.757
1.00
32.52
N


ATOM
7049
CA
ILE
A
463
−36.694
28.843
26.971
1.00
32.32
C


ATOM
7051
CB
ILE
A
463
−37.006
29.724
25.734
1.00
32.47
C


ATOM
7053
CG1
ILE
A
463
−37.152
28.866
24.463
1.00
32.41
C


ATOM
7056
CD1
ILE
A
463
−37.359
29.675
23.191
1.00
32.02
C


ATOM
7060
CG2
ILE
A
463
−35.937
30.810
25.571
1.00
32.44
C


ATOM
7064
C
ILE
A
463
−37.902
28.006
27.309
1.00
32.15
C


ATOM
7065
O
ILE
A
463
−37.886
26.799
27.145
1.00
32.01
O


ATOM
7067
N
SER
A
464
−38.950
28.670
27.780
1.00
32.27
N


ATOM
7068
CA
SER
A
464
−40.163
28.006
28.239
1.00
32.52
C


ATOM
7070
CB
SER
A
464
−40.964
28.974
29.110
1.00
32.65
C


ATOM
7073
OG
SER
A
464
−41.112
30.224
28.457
1.00
32.69
O


ATOM
7075
C
SER
A
464
−41.036
27.522
27.079
1.00
32.53
C


ATOM
7076
O
SER
A
464
−40.968
28.069
25.986
1.00
32.62
O


ATOM
7078
N
GLU
A
465
−41.857
26.500
27.332
1.00
32.47
N


ATOM
7079
CA
GLU
A
465
−42.836
26.010
26.353
1.00
32.33
C


ATOM
7081
CB
GLU
A
465
−43.689
24.868
26.935
1.00
32.34
C


ATOM
7084
CG
GLU
A
465
−44.756
24.312
25.974
1.00
32.37
C


ATOM
7087
CD
GLU
A
465
−45.675
23.279
26.611
1.00
32.47
C


ATOM
7088
OE1
GLU
A
465
−45.297
22.659
27.630
1.00
31.63
O


ATOM
7089
OE2
GLU
A
465
−46.785
23.086
26.074
1.00
32.77
O


ATOM
7090
C
GLU
A
465
−43.758
27.124
25.864
1.00
32.33
C


ATOM
7091
O
GLU
A
465
−44.078
27.183
24.676
1.00
32.60
O


ATOM
7093
N
GLU
A
466
−44.196
28.001
26.767
1.00
32.08
N


ATOM
7094
CA
GLU
A
466
−45.085
29.097
26.380
1.00
31.79
C


ATOM
7096
CB
GLU
A
466
−45.606
29.832
27.624
1.00
31.91
C


ATOM
7099
CG
GLU
A
466
−46.503
31.041
27.304
1.00
32.82
C


ATOM
7102
CD
GLU
A
466
−47.426
31.472
28.452
1.00
33.67
C


ATOM
7103
OE1
GLU
A
466
−47.519
30.756
29.478
1.00
35.09
O


ATOM
7104
OE2
GLU
A
466
−48.077
32.533
28.312
1.00
33.12
O


ATOM
7105
C
GLU
A
466
−44.392
30.052
25.386
1.00
31.13
C


ATOM
7106
O
GLU
A
466
−45.032
30.603
24.492
1.00
30.87
O


ATOM
7108
N
LEU
A
467
−43.077
30.194
25.531
1.00
30.57
N


ATOM
7109
CA
LEU
A
467
−42.265
31.102
24.708
1.00
30.18
C


ATOM
7111
CB
LEU
A
467
−41.055
31.604
25.524
1.00
30.38
C


ATOM
7114
CG
LEU
A
467
−40.674
33.087
25.463
1.00
30.39
C


ATOM
7116
CD1
LEU
A
467
−41.608
33.900
26.362
1.00
30.23
C


ATOM
7120
CD2
LEU
A
467
−39.214
33.308
25.872
1.00
30.31
C


ATOM
7124
C
LEU
A
467
−41.773
30.401
23.439
1.00
29.47
C


ATOM
7125
O
LEU
A
467
−41.660
31.010
22.382
1.00
29.17
O


ATOM
7127
N
ALA
A
468
−41.445
29.121
23.562
1.00
28.93
N


ATOM
7128
CA
ALA
A
468
−41.095
28.310
22.404
1.00
28.45
C


ATOM
7130
CB
ALA
A
468
−40.613
26.935
22.830
1.00
28.10
C


ATOM
7134
C
ALA
A
468
−42.306
28.193
21.494
1.00
28.08
C


ATOM
7135
O
ALA
A
468
−42.179
28.321
20.285
1.00
28.24
O


ATOM
7137
N
THR
A
469
−43.479
27.964
22.080
1.00
27.58
N


ATOM
7138
CA
THR
A
469
−44.719
27.870
21.322
1.00
27.26
C


ATOM
7140
CB
THR
A
469
−45.928
27.710
22.261
1.00
27.28
C


ATOM
7142
OG1
THR
A
469
−46.063
26.330
22.627
1.00
27.55
O


ATOM
7144
CG2
THR
A
469
−47.222
28.185
21.597
1.00
27.34
C


ATOM
7148
C
THR
A
469
−44.916
29.099
20.449
1.00
27.10
C


ATOM
7149
O
THR
A
469
−45.176
28.989
19.250
1.00
27.11
O


ATOM
7151
N
GLU
A
470
−44.774
30.267
21.061
1.00
26.88
N


ATOM
7152
CA
GLU
A
470
−44.944
31.542
20.376
1.00
26.84
C


ATOM
7154
CB
GLU
A
470
−44.832
32.663
21.400
1.00
27.07
C


ATOM
7157
CG
GLU
A
470
−45.357
34.003
20.958
1.00
28.47
C


ATOM
7160
CD
GLU
A
470
−45.140
35.087
22.008
1.00
30.51
C


ATOM
7161
OE1
GLU
A
470
−45.029
34.775
23.221
1.00
31.53
O


ATOM
7162
OE2
GLU
A
470
−45.083
36.269
21.608
1.00
32.59
O


ATOM
7163
C
GLU
A
470
−43.926
31.750
19.239
1.00
26.48
C


ATOM
7164
O
GLU
A
470
−44.261
32.296
18.184
1.00
26.29
O


ATOM
7166
N
SER
A
471
−42.686
31.316
19.452
1.00
26.12
N


ATOM
7167
CA
SER
A
471
−41.660
31.386
18.408
1.00
25.85
C


ATOM
7169
CB
SER
A
471
−40.296
30.933
18.941
1.00
25.96
C


ATOM
7172
OG
SER
A
471
−39.656
31.939
19.711
1.00
26.53
O


ATOM
7174
C
SER
A
471
−42.036
30.529
17.208
1.00
25.35
C


ATOM
7175
O
SER
A
471
−41.761
30.899
16.067
1.00
25.43
O


ATOM
7177
N
VAL
A
472
−42.653
29.381
17.482
1.00
24.95
N


ATOM
7178
CA
VAL
A
472
−43.097
28.451
16.438
1.00
24.65
C


ATOM
7180
CB
VAL
A
472
−43.485
27.055
17.024
1.00
24.57
C


ATOM
7182
CG1
VAL
A
472
−44.257
26.222
16.016
1.00
23.23
C


ATOM
7186
CG2
VAL
A
472
−42.232
26.309
17.494
1.00
23.83
C


ATOM
7190
C
VAL
A
472
−44.252
29.039
15.631
1.00
24.71
C


ATOM
7191
O
VAL
A
472
−44.369
28.776
14.440
1.00
24.63
O


ATOM
7193
N
MET
A
473
−45.073
29.866
16.269
1.00
24.79
N


ATOM
7194
CA
MET
A
473
−46.143
30.591
15.565
1.00
24.94
C


ATOM
7196
CB
MET
A
473
−47.059
31.266
16.576
1.00
25.11
C


ATOM
7199
CG
MET
A
473
−47.683
30.335
17.560
1.00
25.11
C


ATOM
7202
SD
MET
A
473
−48.967
29.391
16.780
1.00
25.35
S


ATOM
7203
CE
MET
A
473
−50.064
29.163
18.189
1.00
26.11
C


ATOM
7207
C
MET
A
473
−45.613
31.672
14.605
1.00
24.86
C


ATOM
7208
O
MET
A
473
−46.132
31.849
13.513
1.00
24.57
O


ATOM
7210
N
ASN
A
474
−44.589
32.405
15.032
1.00
24.89
N


ATOM
7211
CA
ASN
A
474
−43.951
33.383
14.166
1.00
25.00
C


ATOM
7213
CB
ASN
A
474
−43.009
34.278
14.966
1.00
25.12
C


ATOM
7216
CG
ASN
A
474
−43.700
34.966
16.127
1.00
25.77
C


ATOM
7217
OD1
ASN
A
474
−43.058
35.312
17.114
1.00
26.96
O


ATOM
7218
ND2
ASN
A
474
−45.015
35.163
16.021
1.00
26.31
N


ATOM
7221
C
ASN
A
474
−43.195
32.707
13.024
1.00
24.84
C


ATOM
7222
O
ASN
A
474
−42.979
33.313
11.973
1.00
25.07
O


ATOM
7224
N
LEU
A
475
−42.792
31.455
13.227
1.00
24.46
N


ATOM
7225
CA
LEU
A
475
−42.218
30.671
12.142
1.00
24.06
C


ATOM
7227
CB
LEU
A
475
−41.596
29.367
12.653
1.00
24.27
C


ATOM
7230
CG
LEU
A
475
−40.307
29.072
11.886
1.00
25.08
C


ATOM
7232
CD1
LEU
A
475
−39.207
30.000
12.419
1.00
25.89
C


ATOM
7236
CD2
LEU
A
475
−39.887
27.622
11.985
1.00
25.33
C


ATOM
7240
C
LEU
A
475
−43.278
30.365
11.085
1.00
23.26
C


ATOM
7241
O
LEU
A
475
−43.018
30.479
9.884
1.00
22.90
O


ATOM
7243
N
ILE
A
476
−44.471
29.978
11.534
1.00
22.53
N


ATOM
7244
CA
ILE
A
476
−45.544
29.632
10.606
1.00
21.93
C


ATOM
7246
CB
ILE
A
476
−46.773
29.017
11.308
1.00
21.58
C


ATOM
7248
CG1
ILE
A
476
−46.499
27.550
11.664
1.00
21.32
C


ATOM
7251
CD1
ILE
A
476
−47.552
26.902
12.598
1.00
20.33
C


ATOM
7255
CG2
ILE
A
476
−47.994
29.104
10.428
1.00
20.46
C


ATOM
7259
C
ILE
A
476
−45.926
30.887
9.853
1.00
22.07
C


ATOM
7260
O
ILE
A
476
−46.007
30.869
8.626
1.00
22.47
O


ATOM
7262
N
ASP
A
477
−46.122
31.982
10.584
1.00
21.81
N


ATOM
7263
CA
ASP
A
477
−46.483
33.245
9.970
1.00
21.56
C


ATOM
7265
CB
ASP
A
477
−46.643
34.337
11.032
1.00
21.89
C


ATOM
7268
CG
ASP
A
477
−47.962
34.217
11.817
1.00
23.39
C


ATOM
7269
OD1
ASP
A
477
−48.854
33.441
11.375
1.00
24.41
O


ATOM
7270
OD2
ASP
A
477
−48.103
34.902
12.875
1.00
23.80
O


ATOM
7271
C
ASP
A
477
−45.425
33.626
8.947
1.00
21.06
C


ATOM
7272
O
ASP
A
477
−45.759
33.913
7.795
1.00
20.78
O


ATOM
7274
N
GLU
A
478
−44.156
33.591
9.361
1.00
20.63
N


ATOM
7275
CA
GLU
A
478
−43.027
33.919
8.467
1.00
20.45
C


ATOM
7277
CB
GLU
A
478
−41.680
33.793
9.200
1.00
20.66
C


ATOM
7280
CG
GLU
A
478
−40.422
34.044
8.339
1.00
22.27
C


ATOM
7283
CD
GLU
A
478
−39.107
33.579
9.018
1.00
24.72
C


ATOM
7284
OE1
GLU
A
478
−38.553
34.367
9.817
1.00
26.19
O


ATOM
7285
OE2
GLU
A
478
−38.619
32.442
8.745
1.00
25.66
O


ATOM
7286
C
GLU
A
478
−43.041
33.042
7.217
1.00
19.54
C


ATOM
7287
O
GLU
A
478
−42.879
33.544
6.102
1.00
19.02
O


ATOM
7289
N
THR
A
479
−43.250
31.743
7.413
1.00
18.76
N


ATOM
7290
CA
THR
A
479
−43.358
30.808
6.294
1.00
18.45
C


ATOM
7292
CB
THR
A
479
−43.503
29.339
6.774
1.00
18.26
C


ATOM
7294
OG1
THR
A
479
−42.334
28.955
7.494
1.00
17.30
O


ATOM
7296
CG2
THR
A
479
−43.669
28.394
5.592
1.00
17.93
C


ATOM
7300
C
THR
A
479
−44.504
31.173
5.314
1.00
18.39
C


ATOM
7301
O
THR
A
479
−44.280
31.200
4.103
1.00
18.58
O


ATOM
7303
N
TRP
A
480
−45.709
31.457
5.818
1.00
17.85
N


ATOM
7304
CA
TRP
A
480
−46.801
31.909
4.942
1.00
17.44
C


ATOM
7306
CB
TRP
A
480
−48.086
32.225
5.728
1.00
17.67
C


ATOM
7309
CG
TRP
A
480
−48.969
31.042
5.888
1.00
17.09
C


ATOM
7310
CD1
TRP
A
480
−48.996
30.195
6.936
1.00
16.76
C


ATOM
7312
NE1
TRP
A
480
−49.914
29.216
6.724
1.00
17.08
N


ATOM
7314
CE2
TRP
A
480
−50.501
29.410
5.507
1.00
17.86
C


ATOM
7315
CD2
TRP
A
480
−49.930
30.559
4.954
1.00
17.15
C


ATOM
7316
CE3
TRP
A
480
−50.356
30.980
3.696
1.00
18.31
C


ATOM
7318
CZ3
TRP
A
480
−51.337
30.244
3.039
1.00
19.10
C


ATOM
7320
CH2
TRP
A
480
−51.897
29.107
3.622
1.00
18.83
C


ATOM
7322
CZ2
TRP
A
480
−51.493
28.673
4.855
1.00
18.86
C


ATOM
7324
C
TRP
A
480
−46.426
33.124
4.097
1.00
17.10
C


ATOM
7325
O
TRP
A
480
−46.824
33.215
2.943
1.00
17.22
O


ATOM
7327
N
LYS
A
481
−45.675
34.062
4.659
1.00
16.60
N


ATOM
7328
CA
LYS
A
481
−45.275
35.215
3.886
1.00
16.23
C


ATOM
7330
CB
LYS
A
481
−44.566
36.243
4.747
1.00
16.42
C


ATOM
7333
CG
LYS
A
481
−45.417
36.954
5.769
1.00
16.42
C


ATOM
7336
CD
LYS
A
481
−44.555
37.979
6.505
1.00
16.40
C


ATOM
7339
CE
LYS
A
481
−45.199
38.504
7.782
1.00
16.66
C


ATOM
7342
NZ
LYS
A
481
−44.184
38.717
8.846
1.00
16.72
N


ATOM
7346
C
LYS
A
481
−44.357
34.782
2.767
1.00
15.94
C


ATOM
7347
O
LYS
A
481
−44.451
35.291
1.674
1.00
16.09
O


ATOM
7349
N
LYS
A
482
−43.461
33.849
3.034
1.00
15.98
N


ATOM
7350
CA
LYS
A
482
−42.559
33.369
1.996
1.00
16.30
C


ATOM
7352
CB
LYS
A
482
−41.409
32.563
2.594
1.00
16.51
C


ATOM
7355
CG
LYS
A
482
−40.354
33.445
3.286
1.00
17.46
C


ATOM
7358
CD
LYS
A
482
−39.486
32.647
4.241
1.00
18.48
C


ATOM
7361
CE
LYS
A
482
−38.625
33.548
5.096
1.00
18.94
C


ATOM
7364
NZ
LYS
A
482
−37.865
32.773
6.129
1.00
20.40
N


ATOM
7368
C
LYS
A
482
−43.296
32.561
.938
1.00
16.40
C


ATOM
7369
O
LYS
A
482
−42.941
32.614
−.229
1.00
16.85
O


ATOM
7371
N
MET
A
483
−44.328
31.824
1.332
1.00
16.57
N


ATOM
7372
CA
MET
A
483
−45.149
31.100
.366
1.00
16.67
C


ATOM
7374
CB
MET
A
483
−46.128
30.156
1.057
1.00
16.55
C


ATOM
7377
CG
MET
A
483
−45.496
28.923
1.675
1.00
16.50
C


ATOM
7380
SD
MET
A
483
−46.715
27.684
2.194
1.00
17.39
S


ATOM
7381
CE
MET
A
483
−47.937
28.704
3.025
1.00
17.27
C


ATOM
7385
C
MET
A
483
−45.928
32.069
−.495
1.00
17.06
C


ATOM
7386
O
MET
A
483
−46.027
31.867
−1.688
1.00
17.17
O


ATOM
7388
N
ASN
A
484
−46.483
33.110
.118
1.00
17.72
N


ATOM
7389
CA
ASN
A
484
−47.305
34.093
−.587
1.00
18.35
C


ATOM
7391
CB
ASN
A
484
−47.861
35.129
.397
1.00
18.26
C


ATOM
7394
CG
ASN
A
484
−48.994
34.601
1.248
1.00
17.35
C


ATOM
7395
OD1
ASN
A
484
−49.597
33.577
.934
1.00
17.54
O


ATOM
7396
ND2
ASN
A
484
−49.304
35.317
2.331
1.00
14.10
N


ATOM
7399
C
ASN
A
484
−46.541
34.839
−1.677
1.00
19.51
C


ATOM
7400
O
ASN
A
484
−47.123
35.250
−2.685
1.00
19.49
O


ATOM
7402
N
LYS
A
485
−45.243
35.035
−1.463
1.00
20.86
N


ATOM
7403
CA
LYS
A
485
−44.394
35.681
−2.456
1.00
22.12
C


ATOM
7405
CB
LYS
A
485
−43.070
36.092
−1.823
1.00
22.08
C


ATOM
7408
CG
LYS
A
485
−42.229
37.060
−2.641
1.00
23.09
C


ATOM
7411
CD
LYS
A
485
−40.783
37.072
−2.109
1.00
25.38
C


ATOM
7414
CE
LYS
A
485
−40.098
38.460
−2.135
1.00
26.12
C


ATOM
7417
NZ
LYS
A
485
−38.943
38.555
−3.086
1.00
26.61
N


ATOM
7421
C
LYS
A
485
−44.164
34.743
−3.656
1.00
23.36
C


ATOM
7422
O
LYS
A
485
−44.120
35.191
−4.802
1.00
23.00
O


ATOM
7424
N
GLU
A
486
−44.027
33.442
−3.400
1.00
24.99
N


ATOM
7425
CA
GLU
A
486
−43.902
32.479
−4.491
1.00
26.35
C


ATOM
7427
CB
GLU
A
486
−43.627
31.061
−3.978
1.00
26.55
C


ATOM
7430
CG
GLU
A
486
−42.985
30.130
−5.033
1.00
28.29
C


ATOM
7433
CD
GLU
A
486
−41.466
30.324
−5.191
1.00
31.11
C


ATOM
7434
OE1
GLU
A
486
−40.981
30.466
−6.341
1.00
32.07
O


ATOM
7435
OE2
GLU
A
486
−40.744
30.335
−4.159
1.00
33.04
O


ATOM
7436
C
GLU
A
486
−45.157
32.501
−5.362
1.00
27.13
C


ATOM
7437
O
GLU
A
486
−45.064
32.618
−6.583
1.00
27.57
O


ATOM
7439
N
LYS
A
487
−46.325
32.422
−4.739
1.00
28.12
N


ATOM
7440
CA
LYS
A
487
−47.582
32.378
−5.485
1.00
29.12
C


ATOM
7442
CB
LYS
A
487
−48.788
32.260
−4.542
1.00
29.21
C


ATOM
7445
CG
LYS
A
487
−50.167
32.296
−5.235
1.00
28.57
C


ATOM
7448
CD
LYS
A
487
−50.403
31.040
−6.036
1.00
27.91
C


ATOM
7451
CE
LYS
A
487
−51.615
31.141
−6.936
1.00
28.38
C


ATOM
7454
NZ
LYS
A
487
−52.874
31.460
−6.211
1.00
28.03
N


ATOM
7458
C
LYS
A
487
−47.756
33.603
−6.352
1.00
30.18
C


ATOM
7459
O
LYS
A
487
−48.308
33.514
−7.446
1.00
30.21
O


ATOM
7461
N
LEU
A
488
−47.295
34.743
−5.844
1.00
31.64
N


ATOM
7462
CA
LEU
A
488
−47.422
36.026
−6.532
1.00
32.65
C


ATOM
7464
CB
LEU
A
488
−47.426
37.144
−5.494
1.00
32.43
C


ATOM
7467
CG
LEU
A
488
−48.091
38.446
−5.907
1.00
32.50
C


ATOM
7469
CD1
LEU
A
488
−49.593
38.288
−5.943
1.00
32.66
C


ATOM
7473
CD2
LEU
A
488
−47.707
39.554
−4.943
1.00
33.38
C


ATOM
7477
C
LEU
A
488
−46.286
36.242
−7.542
1.00
33.99
C


ATOM
7478
O
LEU
A
488
−46.536
36.539
−8.705
1.00
33.72
O


ATOM
7480
N
GLY
A
489
−45.046
36.045
−7.091
1.00
35.85
N


ATOM
7481
CA
GLY
A
489
−43.845
36.361
−7.874
1.00
37.37
C


ATOM
7484
C
GLY
A
489
−43.278
35.221
−8.709
1.00
38.76
C


ATOM
7485
O
GLY
A
489
−42.414
34.467
−8.250
1.00
39.06
O


ATOM
7487
N
GLY
A
490
−43.760
35.119
−9.947
1.00
40.26
N


ATOM
7488
CA
GLY
A
490
−43.285
34.130
−10.927
1.00
41.03
C


ATOM
7491
C
GLY
A
490
−42.193
33.194
−10.436
1.00
41.61
C


ATOM
7492
O
GLY
A
490
−41.010
33.547
−10.423
1.00
41.62
O


ATOM
7494
N
SER
A
491
−42.590
31.996
−10.023
1.00
42.17
N


ATOM
7495
CA
SER
A
491
−41.619
30.962
−9.662
1.00
42.54
C


ATOM
7497
CB
SER
A
491
−42.279
29.909
−8.741
1.00
42.63
C


ATOM
7500
OG
SER
A
491
−43.451
29.345
−9.294
1.00
42.44
O


ATOM
7502
C
SER
A
491
−41.032
30.349
−10.952
1.00
42.50
C


ATOM
7503
O
SER
A
491
−41.117
30.961
−12.030
1.00
42.57
O


ATOM
7505
N
LEU
A
492
−40.405
29.177
−10.843
1.00
42.18
N


ATOM
7506
CA
LEU
A
492
−40.185
28.333
−12.021
1.00
41.96
C


ATOM
7508
CB
LEU
A
492
−39.110
27.270
−11.782
1.00
42.39
C


ATOM
7511
CG
LEU
A
492
−37.695
27.671
−11.353
1.00
44.21
C


ATOM
7513
CD1
LEU
A
492
−36.783
26.438
−11.498
1.00
45.26
C


ATOM
7517
CD2
LEU
A
492
−37.120
28.896
−12.129
1.00
45.51
C


ATOM
7521
C
LEU
A
492
−41.479
27.610
−12.356
1.00
40.88
C


ATOM
7522
O
LEU
A
492
−41.713
27.244
−13.504
1.00
40.78
O


ATOM
7524
N
PHE
A
493
−42.308
27.409
−11.339
1.00
39.61
N


ATOM
7525
CA
PHE
A
493
−43.500
26.595
−11.460
1.00
38.85
C


ATOM
7527
CB
PHE
A
493
−43.783
25.911
−10.124
1.00
38.62
C


ATOM
7530
CG
PHE
A
493
−42.725
24.937
−9.713
1.00
37.16
C


ATOM
7531
CD1
PHE
A
493
−42.901
23.587
−9.916
1.00
35.26
C


ATOM
7533
CE1
PHE
A
493
−41.934
22.698
−9.546
1.00
34.86
C


ATOM
7535
CZ
PHE
A
493
−40.769
23.143
−8.966
1.00
35.09
C


ATOM
7537
CE2
PHE
A
493
−40.574
24.482
−8.759
1.00
35.45
C


ATOM
7539
CD2
PHE
A
493
−41.549
25.374
−9.130
1.00
36.36
C


ATOM
7541
C
PHE
A
493
−44.714
27.412
−11.890
1.00
38.62
C


ATOM
7542
O
PHE
A
493
−44.756
28.621
−11.702
1.00
38.86
O


ATOM
7544
N
ALA
A
494
−45.698
26.731
−12.469
1.00
38.32
N


ATOM
7545
CA
ALA
A
494
−46.977
27.337
−12.843
1.00
38.09
C


ATOM
7547
CB
ALA
A
494
−47.658
26.486
−13.906
1.00
38.17
C


ATOM
7551
C
ALA
A
494
−47.891
27.479
−11.623
1.00
37.76
C


ATOM
7552
O
ALA
A
494
−48.039
26.541
−10.845
1.00
38.15
O


ATOM
7554
N
LYS
A
495
−48.541
28.629
−11.484
1.00
37.15
N


ATOM
7555
CA
LYS
A
495
−49.321
28.947
−10.270
1.00
36.53
C


ATOM
7557
CB
LYS
A
495
−50.116
30.258
−10.477
1.00
36.90
C


ATOM
7560
CG
LYS
A
495
−49.235
31.524
−10.582
1.00
37.72
C


ATOM
7563
CD
LYS
A
495
−50.061
32.826
−10.585
1.00
38.87
C


ATOM
7566
CE
LYS
A
495
−49.140
34.065
−10.665
1.00
39.78
C


ATOM
7569
NZ
LYS
A
495
−49.786
35.369
−10.282
1.00
39.78
N


ATOM
7573
C
LYS
A
495
−50.238
27.809
−9.724
1.00
35.25
C


ATOM
7574
O
LYS
A
495
−50.261
27.563
−8.523
1.00
34.91
O


ATOM
7576
N
PRO
A
496
−50.978
27.107
−10.599
1.00
33.76
N


ATOM
7577
CA
PRO
A
496
−51.846
26.015
−10.162
1.00
32.74
C


ATOM
7579
CB
PRO
A
496
−52.222
25.340
−11.474
1.00
32.99
C


ATOM
7582
CG
PRO
A
496
−52.337
26.470
−12.385
1.00
33.75
C


ATOM
7585
CD
PRO
A
496
−51.196
27.400
−12.022
1.00
33.83
C


ATOM
7588
C
PRO
A
496
−51.194
24.998
−9.267
1.00
31.24
C


ATOM
7589
O
PRO
A
496
−51.822
24.522
−8.331
1.00
31.54
O


ATOM
7590
N
PHE
A
497
−49.954
24.642
−9.566
1.00
29.40
N


ATOM
7591
CA
PHE
A
497
−49.229
23.726
−8.708
1.00
27.76
C


ATOM
7593
CB
PHE
A
497
−48.162
22.948
−9.474
1.00
27.62
C


ATOM
7596
CG
PHE
A
497
−47.351
22.040
−8.597
1.00
26.33
C


ATOM
7597
CD1
PHE
A
497
−47.944
20.969
−7.974
1.00
24.96
C


ATOM
7599
CE1
PHE
A
497
−47.218
20.151
−7.156
1.00
25.04
C


ATOM
7601
CZ
PHE
A
497
−45.875
20.396
−6.940
1.00
24.79
C


ATOM
7603
CE2
PHE
A
497
−45.274
21.457
−7.549
1.00
24.73
C


ATOM
7605
CD2
PHE
A
497
−46.010
22.282
−8.366
1.00
25.48
C


ATOM
7607
C
PHE
A
497
−48.592
24.407
−7.494
1.00
26.59
C


ATOM
7608
O
PHE
A
497
−48.361
23.738
−6.500
1.00
26.77
O


ATOM
7610
N
VAL
A
498
−48.291
25.704
−7.535
1.00
24.83
N


ATOM
7611
CA
VAL
A
498
−47.869
26.338
−6.289
1.00
23.86
C


ATOM
7613
CB
VAL
A
498
−47.181
27.722
−6.455
1.00
23.47
C


ATOM
7615
CG1
VAL
A
498
−48.163
28.755
−6.759
1.00
24.57
C


ATOM
7619
CG2
VAL
A
498
−46.151
27.690
−7.551
1.00
23.54
C


ATOM
7623
C
VAL
A
498
−49.094
26.396
−5.355
1.00
22.97
C


ATOM
7624
O
VAL
A
498
−48.978
26.130
−4.161
1.00
23.39
O


ATOM
7626
N
GLU
A
499
−50.268
26.687
−5.903
1.00
21.68
N


ATOM
7627
CA
GLU
A
499
−51.473
26.719
−5.106
1.00
20.84
C


ATOM
7629
CB
GLU
A
499
−52.677
27.136
−5.930
1.00
20.99
C


ATOM
7632
CG
GLU
A
499
−53.957
27.352
−5.099
1.00
21.25
C


ATOM
7635
CD
GLU
A
499
−53.930
28.630
−4.265
1.00
21.77
C


ATOM
7636
OE1
GLU
A
499
−52.982
29.448
−4.383
1.00
20.74
O


ATOM
7637
OE2
GLU
A
499
−54.876
28.812
−3.476
1.00
22.76
O


ATOM
7638
C
GLU
A
499
−51.766
25.371
−4.497
1.00
20.35
C


ATOM
7639
O
GLU
A
499
−52.137
25.308
−3.318
1.00
20.78
O


ATOM
7641
N
THR
A
500
−51.629
24.286
−5.264
1.00
19.40
N


ATOM
7642
CA
THR
A
500
−51.894
22.970
−4.667
1.00
18.94
C


ATOM
7644
CB
THR
A
500
−51.993
21.782
−5.683
1.00
18.96
C


ATOM
7646
OG1
THR
A
500
−50.708
21.475
−6.202
1.00
19.44
O


ATOM
7648
CG2
THR
A
500
−52.991
22.071
−6.835
1.00
18.75
C


ATOM
7652
C
THR
A
500
−50.895
22.674
−3.526
1.00
18.20
C


ATOM
7653
O
THR
A
500
−51.280
22.100
−2.519
1.00
17.68
O


ATOM
7655
N
ALA
A
501
−49.646
23.119
−3.666
1.00
17.49
N


ATOM
7656
CA
ALA
A
501
−48.663
23.021
−2.593
1.00
17.18
C


ATOM
7658
CB
ALA
A
501
−47.348
23.563
−3.051
1.00
16.98
C


ATOM
7662
C
ALA
A
501
−49.120
23.770
−1.338
1.00
17.38
C


ATOM
7663
O
ALA
A
501
−49.098
23.219
−.230
1.00
17.45
O


ATOM
7665
N
ILE
A
502
−49.517
25.031
−1.508
1.00
17.29
N


ATOM
7666
CA
ILE
A
502
−49.971
25.857
−.386
1.00
17.07
C


ATOM
7668
CB
ILE
A
502
−50.353
27.304
−.846
1.00
17.09
C


ATOM
7670
CG1
ILE
A
502
−49.092
28.069
−1.286
1.00
17.17
C


ATOM
7673
CD1
ILE
A
502
−49.345
29.345
−2.116
1.00
16.25
C


ATOM
7677
CG2
ILE
A
502
−51.110
28.076
.265
1.00
16.14
C


ATOM
7681
C
ILE
A
502
−51.155
25.168
.285
1.00
17.24
C


ATOM
7682
O
ILE
A
502
−51.265
25.163
1.516
1.00
17.03
O


ATOM
7684
N
ASN
A
503
−52.022
24.559
−.522
1.00
17.32
N


ATOM
7685
CA
ASN
A
503
−53.167
23.823
.022
1.00
17.75
C


ATOM
7687
CB
ASN
A
503
−53.986
23.221
−1.121
1.00
17.70
C


ATOM
7690
CG
ASN
A
503
−54.760
24.261
−1.888
1.00
18.41
C


ATOM
7691
OD1
ASN
A
503
−55.058
25.334
−1.382
1.00
18.94
O


ATOM
7692
ND2
ASN
A
503
−55.107
23.939
−3.119
1.00
20.42
N


ATOM
7695
C
ASN
A
503
−52.803
22.727
1.075
1.00
17.74
C


ATOM
7696
O
ASN
A
503
−53.619
22.387
1.949
1.00
17.96
O


ATOM
7698
N
LEU
A
504
−51.589
22.185
.993
1.00
17.23
N


ATOM
7699
CA
LEU
A
504
−51.105
21.249
1.994
1.00
17.10
C


ATOM
7701
CB
LEU
A
504
−49.745
20.686
1.583
1.00
17.28
C


ATOM
7704
CG
LEU
A
504
−49.213
19.528
2.426
1.00
17.29
C


ATOM
7706
CD1
LEU
A
504
−49.570
18.196
1.762
1.00
17.22
C


ATOM
7710
CD2
LEU
A
504
−47.708
19.686
2.616
1.00
16.83
C


ATOM
7714
C
LEU
A
504
−50.971
21.939
3.347
1.00
16.97
C


ATOM
7715
O
LEU
A
504
−51.237
21.330
4.379
1.00
16.84
O


ATOM
7717
N
ALA
A
505
−50.535
23.201
3.330
1.00
16.90
N


ATOM
7718
CA
ALA
A
505
−50.433
24.015
4.540
1.00
16.68
C


ATOM
7720
CB
ALA
A
505
−49.739
25.309
4.243
1.00
16.54
C


ATOM
7724
C
ALA
A
505
−51.826
24.281
5.074
1.00
16.77
C


ATOM
7725
O
ALA
A
505
−52.087
24.123
6.266
1.00
16.80
O


ATOM
7727
N
ARG
A
506
−52.726
24.664
4.172
1.00
16.82
N


ATOM
7728
CA
ARG
A
506
−54.128
24.875
4.521
1.00
16.84
C


ATOM
7730
CB
ARG
A
506
−54.944
25.274
3.286
1.00
16.86
C


ATOM
7733
CG
ARG
A
506
−54.649
26.661
2.795
1.00
16.27
C


ATOM
7736
CD
ARG
A
506
−55.586
27.090
1.726
1.00
15.26
C


ATOM
7739
NE
ARG
A
506
−55.240
28.436
1.273
1.00
15.31
N


ATOM
7741
CZ
ARG
A
506
−54.744
28.758
.082
1.00
14.28
C


ATOM
7742
NH1
ARG
A
506
−54.519
27.856
−.856
1.00
14.12
N


ATOM
7745
NH2
ARG
A
506
−54.471
30.018
−.176
1.00
15.94
N


ATOM
7748
C
ARG
A
506
−54.732
23.631
5.122
1.00
16.96
C


ATOM
7749
O
ARG
A
506
−55.480
23.707
6.089
1.00
16.71
O


ATOM
7751
N
GLN
A
507
−54.415
22.481
4.542
1.00
17.37
N


ATOM
7752
CA
GLN
A
507
−54.962
21.233
5.048
1.00
17.87
C


ATOM
7754
CB
GLN
A
507
−54.712
20.075
4.087
1.00
17.80
C


ATOM
7757
CG
GLN
A
507
−55.293
18.740
4.571
1.00
17.24
C


ATOM
7760
CD
GLN
A
507
−56.777
18.805
4.860
1.00
16.23
C


ATOM
7761
OE1
GLN
A
507
−57.506
19.585
4.250
1.00
16.66
O


ATOM
7762
NE2
GLN
A
507
−57.233
17.980
5.790
1.00
14.96
N


ATOM
7765
C
GLN
A
507
−54.401
20.890
6.423
1.00
18.28
C


ATOM
7766
O
GLN
A
507
−55.148
20.419
7.287
1.00
18.53
O


ATOM
7768
N
SER
A
508
−53.096
21.113
6.609
1.00
18.50
N


ATOM
7769
CA
SER
A
508
−52.435
20.924
7.908
1.00
18.54
C


ATOM
7771
CB
SER
A
508
−50.979
21.385
7.856
1.00
18.40
C


ATOM
7774
OG
SER
A
508
−50.259
20.694
6.857
1.00
18.51
O


ATOM
7776
C
SER
A
508
−53.147
21.752
8.942
1.00
18.78
C


ATOM
7777
O
SER
A
508
−53.535
21.278
10.006
1.00
18.33
O


ATOM
7779
N
HIS
A
509
−53.324
23.014
8.599
1.00
19.37
N


ATOM
7780
CA
HIS
A
509
−54.014
23.922
9.466
1.00
19.92
C


ATOM
7782
CB
HIS
A
509
−54.126
25.295
8.818
1.00
20.07
C


ATOM
7785
CG
HIS
A
509
−53.000
26.188
9.174
1.00
19.82
C


ATOM
7786
ND1
HIS
A
509
−52.079
26.631
8.255
1.00
19.73
N


ATOM
7788
CE1
HIS
A
509
−51.187
27.383
8.866
1.00
20.25
C


ATOM
7790
NE2
HIS
A
509
−51.485
27.426
10.149
1.00
21.66
N


ATOM
7792
CD2
HIS
A
509
−52.615
26.682
10.368
1.00
20.98
C


ATOM
7794
C
HIS
A
509
−55.389
23.448
9.839
1.00
20.41
C


ATOM
7795
O
HIS
A
509
−55.821
23.694
10.934
1.00
20.22
O


ATOM
7797
N
CYS
A
510
−56.093
22.808
8.918
1.00
21.33
N


ATOM
7798
CA
CYS
A
510
−57.470
22.396
9.189
1.00
22.03
C


ATOM
7800
CB
CYS
A
510
−58.278
22.383
7.894
1.00
21.90
C


ATOM
7803
SG
CYS
A
510
−58.522
24.055
7.270
1.00
21.93
S


ATOM
7805
C
CYS
A
510
−57.527
21.054
9.908
1.00
22.60
C


ATOM
7806
O
CYS
A
510
−58.423
20.822
10.706
1.00
22.23
O


ATOM
7808
N
THR
A
511
−56.553
20.194
9.627
1.00
23.67
N


ATOM
7809
CA
THR
A
511
−56.421
18.920
10.305
1.00
24.81
C


ATOM
7811
CB
THR
A
511
−55.437
18.000
9.565
1.00
24.70
C


ATOM
7813
OG1
THR
A
511
−56.071
17.485
8.394
1.00
24.45
O


ATOM
7815
CG2
THR
A
511
−55.003
16.844
10.448
1.00
24.23
C


ATOM
7819
C
THR
A
511
−55.965
19.086
11.756
1.00
26.22
C


ATOM
7820
O
THR
A
511
−56.685
18.717
12.679
1.00
26.12
O


ATOM
7822
N
TYR
A
512
−54.775
19.643
11.959
1.00
28.08
N


ATOM
7823
CA
TYR
A
512
−54.177
19.657
13.298
1.00
29.63
C


ATOM
7825
CB
TYR
A
512
−52.663
19.783
13.238
1.00
29.67
C


ATOM
7828
CG
TYR
A
512
−52.117
18.579
12.560
1.00
29.56
C


ATOM
7829
CD1
TYR
A
512
−51.965
17.395
13.246
1.00
29.66
C


ATOM
7831
CE1
TYR
A
512
−51.507
16.273
12.614
1.00
30.94
C


ATOM
7833
CZ
TYR
A
512
−51.217
16.326
11.263
1.00
32.04
C


ATOM
7834
OH
TYR
A
512
−50.757
15.209
10.608
1.00
33.78
O


ATOM
7836
CE2
TYR
A
512
−51.390
17.492
10.557
1.00
31.29
C


ATOM
7838
CD2
TYR
A
512
−51.847
18.600
11.205
1.00
30.53
C


ATOM
7840
C
TYR
A
512
−54.812
20.676
14.206
1.00
31.11
C


ATOM
7841
O
TYR
A
512
−55.494
20.277
15.139
1.00
31.34
O


ATOM
7843
N
HIS
A
513
−54.583
21.969
13.966
1.00
32.89
N


ATOM
7844
CA
HIS
A
513
−55.505
23.023
14.442
1.00
34.48
C


ATOM
7846
CB
HIS
A
513
−56.372
23.461
13.225
1.00
35.13
C


ATOM
7849
CG
HIS
A
513
−57.704
24.108
13.529
1.00
37.10
C


ATOM
7850
ND1
HIS
A
513
−58.265
25.040
12.678
1.00
38.69
N


ATOM
7852
CE1
HIS
A
513
−59.442
25.416
13.151
1.00
39.23
C


ATOM
7854
NE2
HIS
A
513
−59.678
24.751
14.268
1.00
38.18
N


ATOM
7856
CD2
HIS
A
513
−58.615
23.915
14.519
1.00
38.17
C


ATOM
7858
C
HIS
A
513
−56.337
22.503
15.610
1.00
35.00
C


ATOM
7859
O
HIS
A
513
−56.302
23.074
16.712
1.00
35.48
O


ATOM
7861
N
ASN
A
514
−57.079
21.419
15.351
1.00
35.23
N


ATOM
7862
CA
ASN
A
514
−57.934
20.768
16.347
1.00
35.46
C


ATOM
7864
CB
ASN
A
514
−58.342
19.343
15.903
1.00
35.39
C


ATOM
7867
CG
ASN
A
514
−59.383
19.345
14.786
1.00
34.01
C


ATOM
7868
OD1
ASN
A
514
−59.336
20.185
13.896
1.00
33.36
O


ATOM
7869
ND2
ASN
A
514
−60.315
18.406
14.833
1.00
31.13
N


ATOM
7872
C
ASN
A
514
−57.464
20.751
17.818
1.00
36.15
C


ATOM
7873
O
ASN
A
514
−56.397
20.211
18.153
1.00
36.13
O


ATOM
7875
N
GLY
A
515
−58.277
21.430
18.640
1.00
36.95
N


ATOM
7876
CA
GLY
A
515
−58.427
21.235
20.079
1.00
37.43
C


ATOM
7879
C
GLY
A
515
−59.724
20.470
20.381
1.00
38.18
C


ATOM
7880
O
GLY
A
515
−59.636
19.418
21.021
1.00
38.57
O


ATOM
7882
N
ASP
A
516
−60.930
20.921
19.969
1.00
38.73
N


ATOM
7883
CA
ASP
A
516
−61.249
22.194
19.293
1.00
39.26
C


ATOM
7885
CB
ASP
A
516
−61.607
21.904
17.845
1.00
39.51
C


ATOM
7888
CG
ASP
A
516
−60.523
22.291
16.894
1.00
42.32
C


ATOM
7889
OD1
ASP
A
516
−59.613
23.084
17.286
1.00
45.56
O


ATOM
7890
OD2
ASP
A
516
−60.578
21.797
15.740
1.00
45.52
O


ATOM
7891
C
ASP
A
516
−62.466
22.913
19.884
1.00
39.38
C


ATOM
7892
O
ASP
A
516
−63.032
22.454
20.880
1.00
39.74
O


ATOM
7894
N
ALA
A
517
−62.850
24.038
19.259
1.00
39.40
N


ATOM
7895
CA
ALA
A
517
−64.140
24.752
19.481
1.00
39.50
C


ATOM
7897
CB
ALA
A
517
−65.117
24.403
18.333
1.00
39.27
C


ATOM
7901
C
ALA
A
517
−64.837
24.566
20.863
1.00
39.90
C


ATOM
7902
O
ALA
A
517
−64.173
24.428
21.899
1.00
39.89
O


ATOM
7904
N
HIS
A
518
−66.175
24.617
20.876
1.00
40.33
N


ATOM
7905
CA
HIS
A
518
−66.973
24.112
22.016
1.00
40.73
C


ATOM
7907
CB
HIS
A
518
−68.091
25.092
22.396
1.00
41.22
C


ATOM
7910
CG
HIS
A
518
−67.578
26.419
22.866
1.00
43.58
C


ATOM
7911
ND1
HIS
A
518
−67.426
26.725
24.207
1.00
45.88
N


ATOM
7913
CE1
HIS
A
518
−66.938
27.950
24.320
1.00
46.42
C


ATOM
7915
NE2
HIS
A
518
−66.760
28.446
23.102
1.00
46.25
N


ATOM
7917
CD2
HIS
A
518
−67.145
27.506
22.174
1.00
45.24
C


ATOM
7919
C
HIS
A
518
−67.526
22.725
21.655
1.00
40.17
C


ATOM
7920
O
HIS
A
518
−68.738
22.501
21.571
1.00
40.02
O


ATOM
7922
N
THR
A
519
−66.581
21.812
21.448
1.00
39.64
N


ATOM
7923
CA
THR
A
519
−66.808
20.466
20.918
1.00
39.16
C


ATOM
7925
CB
THR
A
519
−67.414
20.469
19.453
1.00
39.16
C


ATOM
7927
OG1
THR
A
519
−66.847
21.529
18.665
1.00
39.04
O


ATOM
7929
CG2
THR
A
519
−68.942
20.633
19.480
1.00
38.82
C


ATOM
7933
C
THR
A
519
−65.428
19.771
20.978
1.00
38.85
C


ATOM
7934
O
THR
A
519
−64.391
20.433
20.878
1.00
38.66
O


ATOM
7936
N
SER
A
520
−65.407
18.455
21.170
1.00
38.37
N


ATOM
7937
CA
SER
A
520
−64.160
17.745
21.500
1.00
38.08
C


ATOM
7939
CB
SER
A
520
−64.514
16.320
21.931
1.00
38.06
C


ATOM
7942
OG
SER
A
520
−64.560
15.475
20.805
1.00
38.66
O


ATOM
7944
C
SER
A
520
−63.136
17.771
20.327
1.00
37.86
C


ATOM
7945
O
SER
A
520
−63.420
18.369
19.289
1.00
37.50
O


ATOM
7947
N
PRO
A
521
−61.948
17.122
20.482
1.00
37.98
N


ATOM
7948
CA
PRO
A
521
−60.944
17.170
19.379
1.00
37.92
C


ATOM
7950
CB
PRO
A
521
−59.674
16.527
19.987
1.00
37.87
C


ATOM
7953
CG
PRO
A
521
−60.091
15.919
21.330
1.00
38.28
C


ATOM
7956
CD
PRO
A
521
−61.577
16.137
21.526
1.00
38.13
C


ATOM
7959
C
PRO
A
521
−61.432
16.446
18.110
1.00
37.96
C


ATOM
7960
O
PRO
A
521
−61.769
17.118
17.140
1.00
38.09
O


ATOM
7961
N
ASP
A
522
−61.483
15.108
18.107
1.00
38.07
N


ATOM
7962
CA
ASP
A
522
−62.351
14.393
17.161
1.00
38.14
C


ATOM
7964
CB
ASP
A
522
−62.132
12.877
17.204
1.00
38.31
C


ATOM
7967
CG
ASP
A
522
−60.769
12.456
16.640
1.00
39.45
C


ATOM
7968
OD1
ASP
A
522
−60.160
13.200
15.827
1.00
39.72
O


ATOM
7969
OD2
ASP
A
522
−60.302
11.361
17.025
1.00
41.59
O


ATOM
7970
C
ASP
A
522
−63.760
14.780
17.591
1.00
37.92
C


ATOM
7971
O
ASP
A
522
−63.908
15.608
18.474
1.00
38.01
O


ATOM
7973
N
GLU
A
523
−64.799
14.239
16.974
1.00
37.79
N


ATOM
7974
CA
GLU
A
523
−66.150
14.818
17.121
1.00
37.83
C


ATOM
7976
CB
GLU
A
523
−66.618
14.919
18.591
1.00
37.85
C


ATOM
7979
CG
GLU
A
523
−66.346
13.678
19.483
1.00
39.40
C


ATOM
7982
CD
GLU
A
523
−66.916
13.802
20.931
1.00
41.49
C


ATOM
7983
OE1
GLU
A
523
−67.714
14.737
21.207
1.00
42.81
O


ATOM
7984
OE2
GLU
A
523
−66.566
12.957
21.800
1.00
42.04
O


ATOM
7985
C
GLU
A
523
−66.242
16.203
16.430
1.00
37.51
C


ATOM
7986
O
GLU
A
523
−67.338
16.751
16.283
1.00
37.70
O


ATOM
7988
N
LEU
A
524
−65.097
16.775
16.043
1.00
37.03
N


ATOM
7989
CA
LEU
A
524
−65.044
17.863
15.067
1.00
36.58
C


ATOM
7991
CB
LEU
A
524
−64.174
19.022
15.559
1.00
36.56
C


ATOM
7994
CG
LEU
A
524
−63.963
20.225
14.624
1.00
36.35
C


ATOM
7996
CD1
LEU
A
524
−63.400
21.376
15.418
1.00
35.74
C


ATOM
8000
CD2
LEU
A
524
−65.242
20.681
13.910
1.00
36.53
C


ATOM
8004
C
LEU
A
524
−64.478
17.287
13.782
1.00
36.20
C


ATOM
8005
O
LEU
A
524
−65.106
17.363
12.733
1.00
36.14
O


ATOM
8007
N
THR
A
525
−63.298
16.686
13.869
1.00
35.83
N


ATOM
8008
CA
THR
A
525
−62.756
15.935
12.748
1.00
35.66
C


ATOM
8010
CB
THR
A
525
−61.585
15.015
13.173
1.00
35.48
C


ATOM
8012
OG1
THR
A
525
−60.776
15.678
14.152
1.00
35.55
O


ATOM
8014
CG2
THR
A
525
−60.720
14.641
11.978
1.00
34.73
C


ATOM
8018
C
THR
A
525
−63.875
15.098
12.123
1.00
35.89
C


ATOM
8019
O
THR
A
525
−64.040
15.095
10.909
1.00
35.98
O


ATOM
8021
N
ARG
A
526
−64.667
14.420
12.954
1.00
36.06
N


ATOM
8022
CA
ARG
A
526
−65.739
13.574
12.448
1.00
36.18
C


ATOM
8024
CB
ARG
A
526
−66.340
12.718
13.555
1.00
36.56
C


ATOM
8027
CG
ARG
A
526
−67.416
11.754
13.054
1.00
38.35
C


ATOM
8030
CD
ARG
A
526
−67.781
10.702
14.092
1.00
41.00
C


ATOM
8033
NE
ARG
A
526
−67.740
11.207
15.470
1.00
43.08
N


ATOM
8035
CZ
ARG
A
526
−68.637
12.033
16.024
1.00
44.51
C


ATOM
8036
NH1
ARG
A
526
−69.681
12.500
15.336
1.00
44.46
N


ATOM
8039
NH2
ARG
A
526
−68.476
12.406
17.288
1.00
45.02
N


ATOM
8042
C
ARG
A
526
−66.834
14.384
11.774
1.00
35.68
C


ATOM
8043
O
ARG
A
526
−67.253
14.035
10.675
1.00
35.84
O


ATOM
8045
N
LYS
A
527
−67.309
15.447
12.424
1.00
35.06
N


ATOM
8046
CA
LYS
A
527
−68.285
16.343
11.780
1.00
34.61
C


ATOM
8048
CB
LYS
A
527
−68.680
17.528
12.683
1.00
34.74
C


ATOM
8051
CG
LYS
A
527
−69.818
17.225
13.651
1.00
35.40
C


ATOM
8054
CD
LYS
A
527
−70.301
18.452
14.441
1.00
35.99
C


ATOM
8057
CE
LYS
A
527
−71.280
18.021
15.556
1.00
36.42
C


ATOM
8060
NZ
LYS
A
527
−71.652
19.095
16.530
1.00
36.31
N


ATOM
8064
C
LYS
A
527
−67.723
16.860
10.457
1.00
33.77
C


ATOM
8065
O
LYS
A
527
−68.377
16.763
9.423
1.00
33.75
O


ATOM
8067
N
ARG
A
528
−66.501
17.388
10.499
1.00
32.71
N


ATOM
8068
CA
ARG
A
528
−65.840
17.929
9.310
1.00
31.79
C


ATOM
8070
CB
ARG
A
528
−64.425
18.464
9.644
1.00
31.74
C


ATOM
8073
CG
ARG
A
528
−64.419
19.844
10.330
1.00
30.90
C


ATOM
8076
CD
ARG
A
528
−63.021
20.439
10.527
1.00
29.75
C


ATOM
8079
NE
ARG
A
528
−63.098
21.745
11.190
1.00
29.55
N


ATOM
8081
CZ
ARG
A
528
−62.056
22.513
11.528
1.00
29.96
C


ATOM
8082
NH1
ARG
A
528
−60.801
22.145
11.284
1.00
29.74
N


ATOM
8085
NH2
ARG
A
528
−62.269
23.677
12.130
1.00
30.71
N


ATOM
8088
C
ARG
A
528
−65.796
16.904
8.172
1.00
30.99
C


ATOM
8089
O
ARG
A
528
−66.226
17.206
7.068
1.00
30.97
O


ATOM
8091
N
VAL
A
529
−65.313
15.694
8.450
1.00
30.12
N


ATOM
8092
CA
VAL
A
529
−65.239
14.631
7.438
1.00
29.43
C


ATOM
8094
CB
VAL
A
529
−64.557
13.354
7.972
1.00
29.39
C


ATOM
8096
CG1
VAL
A
529
−64.785
12.187
7.023
1.00
28.89
C


ATOM
8100
CG2
VAL
A
529
−63.062
13.589
8.186
1.00
29.44
C


ATOM
8104
C
VAL
A
529
−66.609
14.243
6.887
1.00
28.95
C


ATOM
8105
O
VAL
A
529
−66.755
14.016
5.690
1.00
29.09
O


ATOM
8107
N
LEU
A
530
−67.615
14.153
7.744
1.00
28.18
N


ATOM
8108
CA
LEU
A
530
−68.956
13.874
7.246
1.00
27.72
C


ATOM
8110
CB
LEU
A
530
−69.971
13.715
8.395
1.00
27.75
C


ATOM
8113
CG
LEU
A
530
−70.334
12.274
8.775
1.00
27.36
C


ATOM
8115
CD1
LEU
A
530
−69.096
11.415
9.050
1.00
26.86
C


ATOM
8119
CD2
LEU
A
530
−71.273
12.279
9.968
1.00
27.14
C


ATOM
8123
C
LEU
A
530
−69.401
14.963
6.256
1.00
27.16
C


ATOM
8124
O
LEU
A
530
−69.861
14.652
5.161
1.00
27.42
O


ATOM
8126
N
SER
A
531
−69.230
16.229
6.635
1.00
26.21
N


ATOM
8127
CA
SER
A
531
−69.730
17.359
5.853
1.00
25.12
C


ATOM
8129
CB
SER
A
531
−69.519
18.657
6.618
1.00
25.01
C


ATOM
8132
OG
SER
A
531
−68.167
18.791
6.985
1.00
23.94
O


ATOM
8134
C
SER
A
531
−69.055
17.476
4.507
1.00
24.48
C


ATOM
8135
O
SER
A
531
−69.687
17.886
3.538
1.00
24.14
O


ATOM
8137
N
VAL
A
532
−67.774
17.115
4.461
1.00
23.89
N


ATOM
8138
CA
VAL
A
532
−66.962
17.226
3.246
1.00
23.53
C


ATOM
8140
CB
VAL
A
532
−65.470
17.437
3.575
1.00
23.09
C


ATOM
8142
CG1
VAL
A
532
−64.633
17.355
2.348
1.00
22.32
C


ATOM
8146
CG2
VAL
A
532
−65.268
18.773
4.198
1.00
23.07
C


ATOM
8150
C
VAL
A
532
−67.100
16.028
2.316
1.00
23.67
C


ATOM
8151
O
VAL
A
532
−67.137
16.209
1.099
1.00
23.58
O


ATOM
8153
N
ILE
A
533
−67.187
14.824
2.886
1.00
23.85
N


ATOM
8154
CA
ILE
A
533
−67.172
13.578
2.107
1.00
24.08
C


ATOM
8156
CB
ILE
A
533
−66.172
12.555
2.691
1.00
24.10
C


ATOM
8158
CG1
ILE
A
533
−64.745
13.079
2.631
1.00
23.47
C


ATOM
8161
CD1
ILE
A
533
−64.178
13.083
1.255
1.00
23.42
C


ATOM
8165
CG2
ILE
A
533
−66.254
11.231
1.932
1.00
24.56
C


ATOM
8169
C
ILE
A
533
−68.522
12.859
1.990
1.00
24.27
C


ATOM
8170
O
ILE
A
533
−68.991
12.620
.887
1.00
24.34
O


ATOM
8172
N
THR
A
534
−69.133
12.487
3.111
1.00
24.57
N


ATOM
8173
CA
THR
A
534
−70.279
11.564
3.072
1.00
24.92
C


ATOM
8175
CB
THR
A
534
−70.200
10.522
4.207
1.00
24.93
C


ATOM
8177
OG1
THR
A
534
−70.491
11.149
5.458
1.00
25.28
O


ATOM
8179
CG2
THR
A
534
−68.804
9.885
4.257
1.00
24.75
C


ATOM
8183
C
THR
A
534
−71.673
12.209
3.083
1.00
25.00
C


ATOM
8184
O
THR
A
534
−72.601
11.646
2.522
1.00
24.94
O


ATOM
8186
N
GLU
A
535
−71.821
13.374
3.708
1.00
25.26
N


ATOM
8187
CA
GLU
A
535
−73.132
14.024
3.833
1.00
25.34
C


ATOM
8189
CB
GLU
A
535
−73.333
14.523
5.255
1.00
25.44
C


ATOM
8192
CG
GLU
A
535
−73.753
13.424
6.196
1.00
25.80
C


ATOM
8195
CD
GLU
A
535
−74.307
13.963
7.468
1.00
25.72
C


ATOM
8196
OE1
GLU
A
535
−75.375
13.487
7.881
1.00
24.50
O


ATOM
8197
OE2
GLU
A
535
−73.678
14.877
8.039
1.00
27.03
O


ATOM
8198
C
GLU
A
535
−73.363
15.179
2.859
1.00
25.26
C


ATOM
8199
O
GLU
A
535
−72.686
16.206
2.939
1.00
25.02
O


ATOM
8201
N
PRO
A
536
−74.351
15.030
1.959
1.00
25.25
N


ATOM
8202
CA
PRO
A
536
−74.655
16.111
1.050
1.00
25.18
C


ATOM
8204
CB
PRO
A
536
−75.661
15.484
.076
1.00
25.15
C


ATOM
8207
CG
PRO
A
536
−75.825
14.077
.476
1.00
24.98
C


ATOM
8210
CD
PRO
A
536
−75.363
13.965
1.868
1.00
25.26
C


ATOM
8213
C
PRO
A
536
−75.289
17.265
1.801
1.00
25.30
C


ATOM
8214
O
PRO
A
536
−75.826
17.070
2.883
1.00
25.57
O


ATOM
8215
N
ILE
A
537
−75.213
18.458
1.230
1.00
25.36
N


ATOM
8216
CA
ILE
A
537
−75.807
19.638
1.827
1.00
25.34
C


ATOM
8218
CB
ILE
A
537
−75.201
20.918
1.221
1.00
25.14
C


ATOM
8220
CG1
ILE
A
537
−73.787
21.131
1.744
1.00
24.32
C


ATOM
8223
CD1
ILE
A
537
−73.228
22.472
1.406
1.00
23.22
C


ATOM
8227
CG2
ILE
A
537
−76.030
22.131
1.569
1.00
25.54
C


ATOM
8231
C
ILE
A
537
−77.313
19.603
1.611
1.00
25.67
C


ATOM
8232
O
ILE
A
537
−77.786
19.132
.579
1.00
25.47
O


ATOM
8234
N
LEU
A
538
−78.071
20.088
2.586
1.00
26.22
N


ATOM
8235
CA
LEU
A
538
−79.514
20.051
2.467
1.00
26.65
C


ATOM
8237
CB
LEU
A
538
−80.213
20.525
3.749
1.00
26.78
C


ATOM
8240
CG
LEU
A
538
−80.162
19.609
4.989
1.00
26.64
C


ATOM
8242
CD1
LEU
A
538
−81.218
20.033
6.007
1.00
26.16
C


ATOM
8246
CD2
LEU
A
538
−80.346
18.138
4.633
1.00
26.12
C


ATOM
8250
C
LEU
A
538
−79.925
20.881
1.262
1.00
27.08
C


ATOM
8253
N
PRO
A
539
−80.786
20.306
.414
1.00
28.03
N


ATOM
8254
CA
PRO
A
539
−81.117
20.855
−.888
1.00
28.44
C


ATOM
8256
CB
PRO
A
539
−82.093
19.828
−1.449
1.00
28.40
C


ATOM
8259
CG
PRO
A
539
−82.793
19.328
−.253
1.00
28.30
C


ATOM
8262
CD
PRO
A
539
−81.720
19.228
.787
1.00
28.11
C


ATOM
8265
C
PRO
A
539
−81.813
22.194
−.804
1.00
28.89
C


ATOM
8266
O
PRO
A
539
−82.396
22.535
.226
1.00
28.97
O


ATOM
8267
N
PHE
A
540
−81.774
22.932
−1.906
1.00
29.32
N


ATOM
8268
CA
PHE
A
540
−82.380
24.245
−1.962
1.00
29.50
C


ATOM
8270
CB
PHE
A
540
−82.146
24.874
−3.326
1.00
29.56
C


ATOM
8273
CG
PHE
A
540
−82.757
26.227
−3.463
1.00
29.75
C


ATOM
8274
CD1
PHE
A
540
−83.874
26.429
−4.255
1.00
29.47
C


ATOM
8276
CE1
PHE
A
540
−84.441
27.681
−4.364
1.00
29.46
C


ATOM
8278
CZ
PHE
A
540
−83.901
28.743
−3.674
1.00
29.81
C


ATOM
8280
CE2
PHE
A
540
−82.791
28.553
−2.877
1.00
30.21
C


ATOM
8282
CD2
PHE
A
540
−82.229
27.300
−2.770
1.00
30.17
C


ATOM
8284
C
PHE
A
540
−83.865
24.129
−1.722
1.00
29.66
C


ATOM
8285
O
PHE
A
540
−84.568
23.551
−2.545
1.00
29.66
O


ATOM
8287
N
GLU
A
541
−84.337
24.673
−.601
1.00
29.90
N


ATOM
8288
CA
GLU
A
541
−85.761
24.605
−.233
1.00
30.06
C


ATOM
8290
CB
GLU
A
541
−85.951
23.724
1.017
1.00
30.15
C


ATOM
8293
CG
GLU
A
541
−87.411
23.361
1.367
1.00
30.64
C


ATOM
8296
CD
GLU
A
541
−88.091
24.351
2.322
1.00
31.35
C


ATOM
8297
OE1
GLU
A
541
−87.468
25.370
2.680
1.00
32.67
O


ATOM
8298
OE2
GLU
A
541
−89.254
24.114
2.719
1.00
30.48
O


ATOM
8299
C
GLU
A
541
−86.315
26.011
−.010
1.00
29.89
C


ATOM
8300
O
GLU
A
541
−86.835
26.636
−.936
1.00
29.71
O


ATOM
8302
N
LEU
B
17
−69.666
−25.325
2.227
1.00
33.20
N


ATOM
8303
CA
LEU
B
17
−69.356
−25.417
.755
1.00
33.49
C


ATOM
8305
CB
LEU
B
17
−70.240
−26.475
.048
1.00
33.44
C


ATOM
8308
CG
LEU
B
17
−70.077
−27.986
.328
1.00
33.60
C


ATOM
8310
CD1
LEU
B
17
−71.285
−28.778
−.217
1.00
32.76
C


ATOM
8314
CD2
LEU
B
17
−68.763
−28.553
−.230
1.00
33.21
C


ATOM
8318
C
LEU
B
17
−69.513
−24.044
.055
1.00
33.54
C


ATOM
8319
O
LEU
B
17
−70.550
−23.380
.195
1.00
33.77
O


ATOM
8323
N
LEU
B
18
−68.481
−23.637
−.696
1.00
33.44
N


ATOM
8324
CA
LEU
B
18
−68.476
−22.362
−1.454
1.00
33.07
C


ATOM
8326
CB
LEU
B
18
−67.029
−21.960
−1.840
1.00
33.21
C


ATOM
8329
CG
LEU
B
18
−66.065
−21.488
−.721
1.00
34.06
C


ATOM
8331
CD1
LEU
B
18
−64.607
−21.351
−1.235
1.00
34.51
C


ATOM
8335
CD2
LEU
B
18
−66.516
−20.164
−.061
1.00
33.43
C


ATOM
8339
C
LEU
B
18
−69.379
−22.400
−2.714
1.00
32.37
C


ATOM
8340
O
LEU
B
18
−69.710
−21.355
−3.274
1.00
32.29
O


ATOM
8342
N
SER
B
19
−69.765
−23.597
−3.153
1.00
31.62
N


ATOM
8343
CA
SER
B
19
−70.711
−23.749
−4.253
1.00
31.16
C


ATOM
8345
CB
SER
B
19
−70.443
−25.067
−5.033
1.00
31.09
C


ATOM
8348
OG
SER
B
19
−70.977
−26.243
−4.421
1.00
29.36
O


ATOM
8350
C
SER
B
19
−72.168
−23.663
−3.759
1.00
31.45
C


ATOM
8351
O
SER
B
19
−73.076
−23.432
−4.551
1.00
31.13
O


ATOM
8353
N
SER
B
20
−72.386
−23.824
−2.451
1.00
31.91
N


ATOM
8354
CA
SER
B
20
−73.749
−23.900
−1.879
1.00
32.34
C


ATOM
8356
CB
SER
B
20
−73.706
−24.122
−.357
1.00
32.35
C


ATOM
8359
OG
SER
B
20
−73.393
−25.473
−.055
1.00
32.34
O


ATOM
8361
C
SER
B
20
−74.601
−22.670
−2.204
1.00
32.69
C


ATOM
8362
O
SER
B
20
−74.072
−21.600
−2.487
1.00
32.89
O


ATOM
8364
N
ASP
B
21
−75.921
−22.831
−2.119
1.00
33.13
N


ATOM
8365
CA
ASP
B
21
−76.874
−21.901
−2.745
1.00
33.40
C


ATOM
8367
CB
ASP
B
21
−78.179
−22.628
−3.045
1.00
33.64
C


ATOM
8370
CG
ASP
B
21
−77.943
−23.883
−3.828
1.00
35.35
C


ATOM
8371
OD1
ASP
B
21
−77.031
−23.866
−4.684
1.00
38.26
O


ATOM
8372
OD2
ASP
B
21
−78.631
−24.890
−3.590
1.00
37.55
O


ATOM
8373
C
ASP
B
21
−77.141
−20.649
−1.940
1.00
33.23
C


ATOM
8374
O
ASP
B
21
−78.039
−20.616
−1.106
1.00
32.91
O


ATOM
8376
N
THR
B
22
−76.348
−19.621
−2.229
1.00
33.46
N


ATOM
8377
CA
THR
B
22
−76.443
−18.307
−1.593
1.00
33.79
C


ATOM
8379
CB
THR
B
22
−76.460
−18.378
−.016
1.00
33.74
C


ATOM
8381
OG1
THR
B
22
−75.426
−19.250
.460
1.00
32.97
O


ATOM
8383
CG2
THR
B
22
−77.805
−18.838
.535
1.00
33.55
C


ATOM
8387
C
THR
B
22
−75.217
−17.479
−2.018
1.00
34.24
C


ATOM
8388
O
THR
B
22
−74.102
−17.988
−1.892
1.00
33.95
O


ATOM
8390
N
ASP
B
23
−75.361
−16.248
−2.543
1.00
34.99
N


ATOM
8391
CA
ASP
B
23
−76.603
−15.620
−3.103
1.00
35.61
C


ATOM
8393
CB
ASP
B
23
−77.242
−16.538
−4.170
1.00
35.51
C


ATOM
8396
CG
ASP
B
23
−76.196
−17.236
−5.034
1.00
36.08
C


ATOM
8397
OD1
ASP
B
23
−75.084
−16.685
−5.194
1.00
35.58
O


ATOM
8398
OD2
ASP
B
23
−76.473
−18.344
−5.541
1.00
37.65
O


ATOM
8399
C
ASP
B
23
−77.662
−15.083
−2.097
1.00
36.10
C


ATOM
8400
O
ASP
B
23
−78.644
−15.770
−1.799
1.00
36.13
O


ATOM
8402
N
GLU
B
24
−77.487
−13.845
−1.616
1.00
36.74
N


ATOM
8403
CA
GLU
B
24
−78.358
−13.316
−.543
1.00
37.59
C


ATOM
8405
CB
GLU
B
24
−77.765
−13.666
.838
1.00
37.88
C


ATOM
8408
CG
GLU
B
24
−77.624
−15.173
1.120
1.00
38.51
C


ATOM
8411
CD
GLU
B
24
−77.314
−15.510
2.588
1.00
39.48
C


ATOM
8412
OE1
GLU
B
24
−77.171
−14.581
3.434
1.00
39.60
O


ATOM
8413
OE2
GLU
B
24
−77.221
−16.725
2.884
1.00
39.51
O


ATOM
8414
C
GLU
B
24
−78.730
−11.809
−.559
1.00
38.11
C


ATOM
8415
O
GLU
B
24
−79.840
−11.453
−.972
1.00
38.20
O


ATOM
8417
N
SER
B
25
−77.825
−10.941
−.089
1.00
38.70
N


ATOM
8418
CA
SER
B
25
−78.192
−9.560
.312
1.00
39.21
C


ATOM
8420
CB
SER
B
25
−76.995
−8.783
.916
1.00
39.29
C


ATOM
8423
OG
SER
B
25
−76.267
−8.037
−.056
1.00
39.40
O


ATOM
8425
C
SER
B
25
−78.853
−8.738
−.802
1.00
39.69
C


ATOM
8426
O
SER
B
25
−78.707
−9.050
−1.995
1.00
39.75
O


ATOM
8428
N
ILE
B
26
−79.544
−7.670
−.378
1.00
40.21
N


ATOM
8429
CA
ILE
B
26
−80.554
−6.962
−1.189
1.00
40.55
C


ATOM
8431
CB
ILE
B
26
−79.925
−6.082
−2.319
1.00
40.66
C


ATOM
8433
CG1
ILE
B
26
−79.083
−4.951
−1.700
1.00
40.73
C


ATOM
8436
CD1
ILE
B
26
−78.596
−3.877
−2.698
1.00
40.68
C


ATOM
8440
CG2
ILE
B
26
−81.018
−5.475
−3.203
1.00
40.88
C


ATOM
8444
C
ILE
B
26
−81.575
−7.999
−1.714
1.00
40.72
C


ATOM
8445
O
ILE
B
26
−81.311
−8.710
−2.690
1.00
40.80
O


ATOM
8447
N
GLU
B
27
−82.729
−8.070
−1.037
1.00
40.86
N


ATOM
8448
CA
GLU
B
27
−83.694
−9.190
−1.160
1.00
40.88
C


ATOM
8450
CB
GLU
B
27
−84.782
−9.060
−.061
1.00
40.95
C


ATOM
8453
CG
GLU
B
27
−84.235
−9.257
1.379
1.00
41.28
C


ATOM
8456
CD
GLU
B
27
−85.123
−8.657
2.484
1.00
41.74
C


ATOM
8457
OE1
GLU
B
27
−85.593
−7.504
2.344
1.00
41.14
O


ATOM
8458
OE2
GLU
B
27
−85.332
−9.337
3.514
1.00
42.36
O


ATOM
8459
C
GLU
B
27
−84.290
−9.355
−2.587
1.00
40.75
C


ATOM
8460
O
GLU
B
27
−83.664
−8.936
−3.569
1.00
40.88
O


ATOM
8462
N
VAL
B
28
−85.461
−9.994
−2.712
1.00
40.40
N


ATOM
8463
CA
VAL
B
28
−86.033
−10.361
−4.027
1.00
39.97
C


ATOM
8465
CB
VAL
B
28
−86.034
−9.157
−5.053
1.00
40.03
C


ATOM
8467
CG1
VAL
B
28
−86.494
−9.600
−6.444
1.00
39.63
C


ATOM
8471
CG2
VAL
B
28
−86.900
−7.994
−4.525
1.00
39.86
C


ATOM
8475
C
VAL
B
28
−85.335
−11.600
−4.625
1.00
39.63
C


ATOM
8476
O
VAL
B
28
−85.858
−12.204
−5.567
1.00
39.63
O


ATOM
8478
N
HIS
B
29
−84.175
−11.980
−4.069
1.00
39.21
N


ATOM
8479
CA
HIS
B
29
−83.431
−13.177
−4.504
1.00
38.81
C


ATOM
8481
CB
HIS
B
29
−81.914
−12.913
−4.665
1.00
38.97
C


ATOM
8484
CG
HIS
B
29
−81.571
−11.721
−5.512
1.00
39.99
C


ATOM
8485
ND1
HIS
B
29
−81.181
−11.825
−6.834
1.00
40.79
N


ATOM
8487
CE1
HIS
B
29
−80.931
−10.615
−7.312
1.00
40.76
C


ATOM
8489
NE2
HIS
B
29
−81.134
−9.734
−6.346
1.00
40.27
N


ATOM
8491
CD2
HIS
B
29
−81.525
−10.400
−5.209
1.00
40.17
C


ATOM
8493
C
HIS
B
29
−83.611
−14.340
−3.523
1.00
38.05
C


ATOM
8494
O
HIS
B
29
−82.782
−15.259
−3.526
1.00
38.02
O


ATOM
8496
N
LYS
B
30
−84.667
−14.321
−2.694
1.00
37.06
N


ATOM
8497
CA
LYS
B
30
−85.107
−15.559
−2.022
1.00
36.33
C


ATOM
8499
CB
LYS
B
30
−86.056
−15.286
−.836
1.00
36.27
C


ATOM
8502
CG
LYS
B
30
−85.352
−14.818
.461
1.00
36.09
C


ATOM
8505
CD
LYS
B
30
−86.355
−14.599
1.612
1.00
36.03
C


ATOM
8508
CE
LYS
B
30
−85.811
−13.659
2.701
1.00
36.24
C


ATOM
8511
NZ
LYS
B
30
−86.802
−13.356
3.789
1.00
35.65
N


ATOM
8515
C
LYS
B
30
−85.710
−16.510
−3.094
1.00
35.69
C


ATOM
8516
O
LYS
B
30
−86.706
−17.205
−2.879
1.00
35.35
O


ATOM
8518
N
ASP
B
31
−85.075
−16.484
−4.268
1.00
35.13
N


ATOM
8519
CA
ASP
B
31
−85.145
−17.519
−5.282
1.00
34.64
C


ATOM
8521
CB
ASP
B
31
−84.592
−16.962
−6.623
1.00
34.60
C


ATOM
8524
CG
ASP
B
31
−84.419
−18.032
−7.725
1.00
34.24
C


ATOM
8525
OD1
ASP
B
31
−84.914
−19.165
−7.570
1.00
34.58
O


ATOM
8526
OD2
ASP
B
31
−83.776
−17.732
−8.761
1.00
31.56
O


ATOM
8527
C
ASP
B
31
−84.304
−18.675
−4.721
1.00
34.31
C


ATOM
8528
O
ASP
B
31
−83.122
−18.838
−5.038
1.00
34.03
O


ATOM
8530
N
LYS
B
32
−84.917
−19.429
−3.813
1.00
33.94
N


ATOM
8531
CA
LYS
B
32
−84.380
−20.710
−3.369
1.00
33.36
C


ATOM
8533
CB
LYS
B
32
−83.772
−20.607
−1.973
1.00
33.45
C


ATOM
8536
CG
LYS
B
32
−82.540
−19.693
−1.951
1.00
33.56
C


ATOM
8539
CD
LYS
B
32
−82.025
−19.455
−.532
1.00
34.17
C


ATOM
8542
CE
LYS
B
32
−81.671
−17.987
−.283
1.00
34.16
C


ATOM
8545
NZ
LYS
B
32
−80.746
−17.437
−1.311
1.00
34.07
N


ATOM
8549
C
LYS
B
32
−85.490
−21.764
−3.499
1.00
32.57
C


ATOM
8550
O
LYS
B
32
−85.908
−22.412
−2.538
1.00
32.10
O


ATOM
8552
N
ALA
B
33
−85.976
−21.859
−4.738
1.00
31.66
N


ATOM
8553
CA
ALA
B
33
−86.576
−23.055
−5.259
1.00
30.94
C


ATOM
8555
CB
ALA
B
33
−87.247
−22.773
−6.597
1.00
30.47
C


ATOM
8559
C
ALA
B
33
−85.442
−24.081
−5.403
1.00
30.55
C


ATOM
8560
O
ALA
B
33
−85.684
−25.230
−5.751
1.00
31.12
O


ATOM
8562
N
LYS
B
34
−84.200
−23.660
−5.161
1.00
29.73
N


ATOM
8563
CA
LYS
B
34
−83.107
−24.585
−4.839
1.00
29.09
C


ATOM
8565
CB
LYS
B
34
−81.776
−23.817
−4.865
1.00
29.30
C


ATOM
8568
CG
LYS
B
34
−80.678
−24.474
−5.726
1.00
29.76
C


ATOM
8571
CD
LYS
B
34
−79.633
−23.458
−6.283
1.00
28.74
C


ATOM
8574
CE
LYS
B
34
−78.417
−24.186
−6.890
1.00
27.56
C


ATOM
8577
NZ
LYS
B
34
−78.745
−25.174
−7.935
1.00
26.16
N


ATOM
8581
C
LYS
B
34
−83.376
−25.320
−3.472
1.00
28.37
C


ATOM
8582
O
LYS
B
34
−83.254
−24.755
−2.374
1.00
27.82
O


ATOM
8584
N
LYS
B
35
−83.635
−26.620
−3.604
1.00
27.53
N


ATOM
8585
CA
LYS
B
35
−84.629
−27.429
−2.851
1.00
26.77
C


ATOM
8587
CB
LYS
B
35
−85.729
−26.602
−2.170
1.00
26.81
C


ATOM
8590
CG
LYS
B
35
−87.089
−26.520
−2.907
1.00
27.05
C


ATOM
8593
CD
LYS
B
35
−88.096
−27.604
−2.452
1.00
27.41
C


ATOM
8596
CE
LYS
B
35
−89.394
−27.636
−3.300
1.00
27.21
C


ATOM
8599
NZ
LYS
B
35
−90.501
−26.792
−2.762
1.00
26.28
N


ATOM
8603
C
LYS
B
35
−85.257
−28.357
−3.920
1.00
26.03
C


ATOM
8604
O
LYS
B
35
−85.853
−29.386
−3.626
1.00
26.00
O


ATOM
8606
N
LEU
B
36
−85.171
−27.909
−5.168
1.00
24.98
N


ATOM
8607
CA
LEU
B
36
−85.095
−28.783
−6.319
1.00
24.02
C


ATOM
8609
CB
LEU
B
36
−85.004
−27.930
−7.594
1.00
23.83
C


ATOM
8612
CG
LEU
B
36
−86.208
−27.103
−8.062
1.00
22.23
C


ATOM
8614
CD1
LEU
B
36
−85.762
−25.873
−8.802
1.00
19.93
C


ATOM
8618
CD2
LEU
B
36
−87.092
−27.930
−8.951
1.00
22.11
C


ATOM
8622
C
LEU
B
36
−83.834
−29.659
−6.185
1.00
23.58
C


ATOM
8623
O
LEU
B
36
−83.840
−30.842
−6.508
1.00
23.38
O


ATOM
8625
N
GLU
B
37
−82.742
−29.046
−5.739
1.00
23.18
N


ATOM
8626
CA
GLU
B
37
−81.510
−29.753
−5.396
1.00
22.88
C


ATOM
8628
CB
GLU
B
37
−80.466
−28.766
−4.855
1.00
22.76
C


ATOM
8631
CG
GLU
B
37
−79.038
−29.297
−4.874
1.00
22.24
C


ATOM
8634
CD
GLU
B
37
−78.085
−28.505
−4.009
1.00
21.06
C


ATOM
8635
OE1
GLU
B
37
−78.368
−27.331
−3.737
1.00
19.65
O


ATOM
8636
OE2
GLU
B
37
−77.037
−29.059
−3.615
1.00
20.62
O


ATOM
8637
C
GLU
B
37
−81.748
−30.856
−4.364
1.00
22.95
C


ATOM
8638
O
GLU
B
37
−81.164
−31.933
−4.458
1.00
22.99
O


ATOM
8640
N
ALA
B
38
−82.591
−30.575
−3.372
1.00
23.02
N


ATOM
8641
CA
ALA
B
38
−82.973
−31.565
−2.364
1.00
22.93
C


ATOM
8643
CB
ALA
B
38
−83.966
−30.959
−1.374
1.00
22.73
C


ATOM
8647
C
ALA
B
38
−83.580
−32.778
−3.044
1.00
23.05
C


ATOM
8648
O
ALA
B
38
−83.170
−33.909
−2.813
1.00
22.52
O


ATOM
8650
N
GLU
B
39
−84.546
−32.506
−3.912
1.00
23.66
N


ATOM
8651
CA
GLU
B
39
−85.273
−33.534
−4.642
1.00
24.08
C


ATOM
8653
CB
GLU
B
39
−86.403
−32.891
−5.453
1.00
24.28
C


ATOM
8656
CG
GLU
B
39
−87.405
−33.869
−6.075
1.00
25.14
C


ATOM
8659
CD
GLU
B
39
−88.773
−33.227
−6.341
1.00
26.43
C


ATOM
8660
OE1
GLU
B
39
−89.232
−32.381
−5.519
1.00
25.90
O


ATOM
8661
OE2
GLU
B
39
−89.387
−33.584
−7.375
1.00
26.99
O


ATOM
8662
C
GLU
B
39
−84.362
−34.359
−5.545
1.00
24.18
C


ATOM
8663
O
GLU
B
39
−84.483
−35.579
−5.575
1.00
24.32
O


ATOM
8665
N
VAL
B
40
−83.450
−33.710
−6.269
1.00
24.43
N


ATOM
8666
CA
VAL
B
40
−82.533
−34.437
−7.153
1.00
24.59
C


ATOM
8668
CB
VAL
B
40
−81.734
−33.496
−8.087
1.00
24.65
C


ATOM
8670
CG1
VAL
B
40
−80.611
−34.258
−8.792
1.00
24.14
C


ATOM
8674
CG2
VAL
B
40
−82.662
−32.852
−9.106
1.00
23.89
C


ATOM
8678
C
VAL
B
40
−81.592
−35.304
−6.328
1.00
24.99
C


ATOM
8679
O
VAL
B
40
−81.265
−36.407
−6.717
1.00
24.77
O


ATOM
8681
N
ARG
B
41
−81.184
−34.811
−5.171
1.00
25.79
N


ATOM
8682
CA
ARG
B
41
−80.387
−35.613
−4.248
1.00
26.75
C


ATOM
8684
CB
ARG
B
41
−80.032
−34.823
−2.998
1.00
27.38
C


ATOM
8687
CG
ARG
B
41
−78.568
−34.777
−2.757
1.00
30.18
C


ATOM
8690
CD
ARG
B
41
−78.281
−34.610
−1.273
1.00
34.43
C


ATOM
8693
NE
ARG
B
41
−76.896
−34.198
−1.016
1.00
37.07
N


ATOM
8695
CZ
ARG
B
41
−76.350
−33.045
−1.416
1.00
38.54
C


ATOM
8696
NH1
ARG
B
41
−75.083
−32.797
−1.112
1.00
40.04
N


ATOM
8699
NH2
ARG
B
41
−77.037
−32.145
−2.128
1.00
38.45
N


ATOM
8702
C
ARG
B
41
−81.081
−36.860
−3.762
1.00
26.60
C


ATOM
8703
O
ARG
B
41
−80.446
−37.894
−3.627
1.00
26.89
O


ATOM
8705
N
ARG
B
42
−82.363
−36.733
−3.423
1.00
26.41
N


ATOM
8706
CA
ARG
B
42
−83.159
−37.870
−3.002
1.00
26.09
C


ATOM
8708
CB
ARG
B
42
−84.588
−37.452
−2.674
1.00
25.99
C


ATOM
8711
CG
ARG
B
42
−85.426
−38.562
−2.049
1.00
25.45
C


ATOM
8714
CD
ARG
B
42
−86.783
−38.078
−1.645
1.00
24.26
C


ATOM
8717
NE
ARG
B
42
−87.549
−37.602
−2.794
1.00
23.73
N


ATOM
8719
CZ
ARG
B
42
−88.547
−36.722
−2.717
1.00
24.51
C


ATOM
8720
NH1
ARG
B
42
−88.912
−36.195
−1.545
1.00
24.99
N


ATOM
8723
NH2
ARG
B
42
−89.185
−36.354
−3.816
1.00
24.46
N


ATOM
8726
C
ARG
B
42
−83.167
−38.894
−4.118
1.00
26.28
C


ATOM
8727
O
ARG
B
42
−82.864
−40.054
−3.888
1.00
26.39
O


ATOM
8729
N
GLU
B
43
−83.476
−38.460
−5.334
1.00
26.52
N


ATOM
8730
CA
GLU
B
43
−83.476
−39.370
−6.479
1.00
27.00
C


ATOM
8732
CB
GLU
B
43
−83.982
−38.676
−7.753
1.00
27.50
C


ATOM
8735
CG
GLU
B
43
−85.469
−38.275
−7.699
1.00
29.78
C


ATOM
8738
CD
GLU
B
43
−86.342
−39.301
−6.974
1.00
32.52
C


ATOM
8739
OE1
GLU
B
43
−86.388
−40.455
−7.445
1.00
35.06
O


ATOM
8740
OE2
GLU
B
43
−86.966
−38.963
−5.936
1.00
33.80
O


ATOM
8741
C
GLU
B
43
−82.147
−40.059
−6.769
1.00
26.37
C


ATOM
8742
O
GLU
B
43
−82.162
−41.126
−7.355
1.00
26.63
O


ATOM
8744
N
ILE
B
44
−81.018
−39.472
−6.373
1.00
25.85
N


ATOM
8745
CA
ILE
B
44
−79.717
−40.126
−6.555
1.00
25.55
C


ATOM
8747
CB
ILE
B
44
−78.547
−39.121
−6.692
1.00
25.26
C


ATOM
8749
CG1
ILE
B
44
−78.706
−38.273
−7.953
1.00
24.47
C


ATOM
8752
CD1
ILE
B
44
−77.690
−37.163
−8.114
1.00
23.21
C


ATOM
8756
CG2
ILE
B
44
−77.207
−39.863
−6.778
1.00
25.35
C


ATOM
8760
C
ILE
B
44
−79.404
−41.109
−5.428
1.00
25.84
C


ATOM
8761
O
ILE
B
44
−78.925
−42.208
−5.698
1.00
25.97
O


ATOM
8763
N
ASN
B
45
−79.663
−40.708
−4.177
1.00
26.25
N


ATOM
8764
CA
ASN
B
45
−79.403
−41.542
−2.983
1.00
26.34
C


ATOM
8766
CB
ASN
B
45
−79.284
−40.681
−1.719
1.00
26.01
C


ATOM
8769
CG
ASN
B
45
−78.072
−39.789
−1.723
1.00
24.65
C


ATOM
8770
OD1
ASN
B
45
−76.957
−40.226
−1.434
1.00
22.47
O


ATOM
8771
ND2
ASN
B
45
−78.288
−38.516
−2.009
1.00
23.36
N


ATOM
8774
C
ASN
B
45
−80.488
−42.586
−2.731
1.00
27.18
C


ATOM
8775
O
ASN
B
45
−80.374
−43.390
−1.806
1.00
27.30
O


ATOM
8777
N
ASN
B
46
−81.553
−42.541
−3.527
1.00
28.17
N


ATOM
8778
CA
ASN
B
46
−82.619
−43.536
−3.489
1.00
29.02
C


ATOM
8780
CB
ASN
B
46
−83.558
−43.289
−4.665
1.00
29.05
C


ATOM
8783
CG
ASN
B
46
−84.615
−44.345
−4.806
1.00
28.82
C


ATOM
8784
OD1
ASN
B
46
−84.861
−45.132
−3.895
1.00
28.14
O


ATOM
8785
ND2
ASN
B
46
−85.255
−44.369
−5.968
1.00
29.67
N


ATOM
8788
C
ASN
B
46
−82.076
−44.958
−3.549
1.00
30.14
C


ATOM
8789
O
ASN
B
46
−81.566
−45.408
−4.581
1.00
30.30
O


ATOM
8791
N
GLU
B
47
−82.218
−45.674
−2.446
1.00
31.36
N


ATOM
8792
CA
GLU
B
47
−81.575
−46.968
−2.295
1.00
32.61
C


ATOM
8794
CB
GLU
B
47
−81.617
−47.430
−.823
1.00
33.09
C


ATOM
8797
CG
GLU
B
47
−80.974
−46.466
.218
1.00
34.19
C


ATOM
8800
CD
GLU
B
47
−81.919
−45.337
.721
1.00
34.57
C


ATOM
8801
OE1
GLU
B
47
−82.911
−45.012
.029
1.00
34.53
O


ATOM
8802
OE2
GLU
B
47
−81.652
−44.762
1.805
1.00
34.30
O


ATOM
8803
C
GLU
B
47
−82.205
−48.046
−3.172
1.00
33.18
C


ATOM
8804
O
GLU
B
47
−81.604
−49.088
−3.370
1.00
33.44
O


ATOM
8806
N
LYS
B
48
−83.408
−47.808
−3.692
1.00
34.11
N


ATOM
8807
CA
LYS
B
48
−84.164
−48.853
−4.396
1.00
34.85
C


ATOM
8809
CB
LYS
B
48
−85.379
−49.254
−3.542
1.00
35.09
C


ATOM
8812
CG
LYS
B
48
−85.003
−49.723
−2.127
1.00
36.00
C


ATOM
8815
CD
LYS
B
48
−86.174
−50.338
−1.350
1.00
37.13
C


ATOM
8818
CE
LYS
B
48
−87.312
−49.340
−1.094
1.00
37.90
C


ATOM
8821
NZ
LYS
B
48
−86.908
−48.161
−.272
1.00
38.31
N


ATOM
8825
C
LYS
B
48
−84.586
−48.487
−5.835
1.00
35.13
C


ATOM
8826
O
LYS
B
48
−85.544
−49.030
−6.361
1.00
34.60
O


ATOM
8828
N
ALA
B
49
−83.858
−47.576
−6.474
1.00
36.09
N


ATOM
8829
CA
ALA
B
49
−84.093
−47.261
−7.886
1.00
36.87
C


ATOM
8831
CB
ALA
B
49
−83.550
−45.880
−8.238
1.00
36.73
C


ATOM
8835
C
ALA
B
49
−83.406
−48.325
−8.714
1.00
37.56
C


ATOM
8836
O
ALA
B
49
−82.315
−48.761
−8.363
1.00
37.62
O


ATOM
8838
N
GLU
B
50
−84.028
−48.754
−9.807
1.00
38.54
N


ATOM
8839
CA
GLU
B
50
−83.399
−49.772
−10.652
1.00
39.40
C


ATOM
8841
CB
GLU
B
50
−84.380
−50.384
−11.673
1.00
39.71
C


ATOM
8844
CG
GLU
B
50
−83.959
−51.808
−12.179
1.00
41.33
C


ATOM
8847
CD
GLU
B
50
−83.733
−52.860
−11.039
1.00
42.88
C


ATOM
8848
OE1
GLU
B
50
−84.683
−53.628
−10.718
1.00
42.94
O


ATOM
8849
OE2
GLU
B
50
−82.603
−52.921
−10.472
1.00
42.96
O


ATOM
8850
C
GLU
B
50
−82.171
−49.163
−11.330
1.00
39.43
C


ATOM
8851
O
GLU
B
50
−82.211
−48.017
−11.791
1.00
39.66
O


ATOM
8853
N
PHE
B
51
−81.084
−49.929
−11.376
1.00
39.32
N


ATOM
8854
CA
PHE
B
51
−79.763
−49.360
−11.636
1.00
39.32
C


ATOM
8856
CB
PHE
B
51
−78.675
−50.429
−11.476
1.00
39.62
C


ATOM
8859
CG
PHE
B
51
−78.470
−50.889
−10.032
1.00
41.82
C


ATOM
8860
CD1
PHE
B
51
−78.093
−49.978
−9.032
1.00
42.93
C


ATOM
8862
CE1
PHE
B
51
−77.892
−50.391
−7.705
1.00
43.30
C


ATOM
8864
CZ
PHE
B
51
−78.059
−51.729
−7.359
1.00
44.33
C


ATOM
8866
CE2
PHE
B
51
−78.430
−52.659
−8.341
1.00
44.75
C


ATOM
8868
CD2
PHE
B
51
−78.633
−52.234
−9.677
1.00
44.06
C


ATOM
8870
C
PHE
B
51
−79.646
−48.625
−12.976
1.00
38.57
C


ATOM
8871
O
PHE
B
51
−79.045
−47.564
−13.046
1.00
38.50
O


ATOM
8873
N
LEU
B
52
−80.262
−49.155
−14.024
1.00
37.99
N


ATOM
8874
CA
LEU
B
52
−80.176
−48.539
−15.351
1.00
37.42
C


ATOM
8876
CB
LEU
B
52
−80.774
−49.476
−16.399
1.00
37.66
C


ATOM
8879
CG
LEU
B
52
−80.502
−49.168
−17.876
1.00
38.39
C


ATOM
8881
CD1
LEU
B
52
−80.171
−50.452
−18.652
1.00
38.46
C


ATOM
8885
CD2
LEU
B
52
−81.703
−48.432
−18.500
1.00
39.23
C


ATOM
8889
C
LEU
B
52
−80.824
−47.143
−15.431
1.00
36.69
C


ATOM
8890
O
LEU
B
52
−80.383
−46.312
−16.224
1.00
36.59
O


ATOM
8892
N
THR
B
53
−81.859
−46.879
−14.625
1.00
35.87
N


ATOM
8893
CA
THR
B
53
−82.409
−45.510
−14.513
1.00
34.91
C


ATOM
8895
CB
THR
B
53
−83.784
−45.421
−13.805
1.00
34.72
C


ATOM
8897
OG1
THR
B
53
−84.724
−46.269
−14.455
1.00
34.04
O


ATOM
8899
CG2
THR
B
53
−84.316
−43.981
−13.855
1.00
34.81
C


ATOM
8903
C
THR
B
53
−81.454
−44.642
−13.722
1.00
34.05
C


ATOM
8904
O
THR
B
53
−81.170
−43.513
−14.121
1.00
34.16
O


ATOM
8906
N
LEU
B
54
−80.987
−45.167
−12.593
1.00
32.84
N


ATOM
8907
CA
LEU
B
54
−80.027
−44.461
−11.772
1.00
32.16
C


ATOM
8909
CB
LEU
B
54
−79.506
−45.354
−10.662
1.00
32.27
C


ATOM
8912
CG
LEU
B
54
−78.521
−44.702
−9.698
1.00
32.56
C


ATOM
8914
CD1
LEU
B
54
−79.275
−43.760
−8.769
1.00
32.38
C


ATOM
8918
CD2
LEU
B
54
−77.755
−45.789
−8.917
1.00
33.29
C


ATOM
8922
C
LEU
B
54
−78.870
−44.006
−12.634
1.00
31.50
C


ATOM
8923
O
LEU
B
54
−78.509
−42.840
−12.607
1.00
31.98
O


ATOM
8925
N
LEU
B
55
−78.297
−44.914
−13.418
1.00
30.43
N


ATOM
8926
CA
LEU
B
55
−77.220
−44.540
−14.333
1.00
29.36
C


ATOM
8928
CB
LEU
B
55
−76.765
−45.733
−15.173
1.00
29.23
C


ATOM
8931
CG
LEU
B
55
−76.157
−46.902
−14.391
1.00
28.60
C


ATOM
8933
CD1
LEU
B
55
−75.697
−48.004
−15.348
1.00
27.60
C


ATOM
8937
CD2
LEU
B
55
−75.020
−46.440
−13.469
1.00
27.20
C


ATOM
8941
C
LEU
B
55
−77.678
−43.406
−15.230
1.00
28.60
C


ATOM
8942
O
LEU
B
55
−77.063
−42.365
−15.267
1.00
28.17
O


ATOM
8944
N
GLU
B
56
−78.786
−43.594
−15.919
1.00
28.23
N


ATOM
8945
CA
GLU
B
56
−79.326
−42.530
−16.759
1.00
28.64
C


ATOM
8947
CB
GLU
B
56
−80.567
−43.031
−17.524
1.00
29.27
C


ATOM
8950
CG
GLU
B
56
−80.229
−43.783
−18.829
1.00
31.73
C


ATOM
8953
CD
GLU
B
56
−81.265
−44.860
−19.206
1.00
35.52
C


ATOM
8954
OE1
GLU
B
56
−82.474
−44.693
−18.889
1.00
36.97
O


ATOM
8955
OE2
GLU
B
56
−80.856
−45.877
−19.825
1.00
37.55
O


ATOM
8956
C
GLU
B
56
−79.632
−41.203
−15.995
1.00
27.77
C


ATOM
8957
O
GLU
B
56
−79.561
−40.103
−16.582
1.00
27.84
O


ATOM
8959
N
LEU
B
57
−79.976
−41.303
−14.710
1.00
26.29
N


ATOM
8960
CA
LEU
B
57
−80.158
−40.120
−13.895
1.00
25.06
C


ATOM
8962
CB
LEU
B
57
−80.724
−40.462
−12.514
1.00
24.83
C


ATOM
8965
CG
LEU
B
57
−80.952
−39.298
−11.544
1.00
23.72
C


ATOM
8967
CD1
LEU
B
57
−81.909
−38.278
−12.117
1.00
21.43
C


ATOM
8971
CD2
LEU
B
57
−81.477
−39.830
−10.227
1.00
22.41
C


ATOM
8975
C
LEU
B
57
−78.801
−39.459
−13.780
1.00
24.40
C


ATOM
8976
O
LEU
B
57
−78.591
−38.373
−14.306
1.00
24.70
O


ATOM
8978
N
ILE
B
58
−77.855
−40.133
−13.144
1.00
23.56
N


ATOM
8979
CA
ILE
B
58
−76.509
−39.574
−12.991
1.00
22.91
C


ATOM
8981
CB
ILE
B
58
−75.454
−40.635
−12.599
1.00
22.49
C


ATOM
8983
CG1
ILE
B
58
−75.753
−41.251
−11.235
1.00
22.09
C


ATOM
8986
CD1
ILE
B
58
−74.936
−42.464
−10.926
1.00
20.88
C


ATOM
8990
CG2
ILE
B
58
−74.103
−39.992
−12.534
1.00
22.63
C


ATOM
8994
C
ILE
B
58
−76.062
−38.927
−14.302
1.00
22.64
C


ATOM
8995
O
ILE
B
58
−75.603
−37.796
−14.315
1.00
22.38
O


ATOM
8997
N
ASP
B
59
−76.228
−39.646
−15.404
1.00
22.56
N


ATOM
8998
CA
ASP
B
59
−75.715
−39.196
−16.684
1.00
22.76
C


ATOM
9000
CB
ASP
B
59
−75.926
−40.269
−17.757
1.00
22.93
C


ATOM
9003
CG
ASP
B
59
−75.274
−39.904
−19.088
1.00
24.58
C


ATOM
9004
OD1
ASP
B
59
−74.157
−39.322
−19.081
1.00
25.70
O


ATOM
9005
OD2
ASP
B
59
−75.897
−40.186
−20.142
1.00
27.51
O


ATOM
9006
C
ASP
B
59
−76.343
−37.863
−17.104
1.00
22.40
C


ATOM
9007
O
ASP
B
59
−75.634
−36.939
−17.520
1.00
22.40
O


ATOM
9009
N
ASN
B
60
−77.662
−37.767
−16.991
1.00
21.91
N


ATOM
9010
CA
ASN
B
60
−78.349
−36.500
−17.217
1.00
21.70
C


ATOM
9012
CB
ASN
B
60
−79.867
−36.674
−17.088
1.00
22.12
C


ATOM
9015
CG
ASN
B
60
−80.477
−37.390
−18.268
1.00
22.76
C


ATOM
9016
OD1
ASN
B
60
−80.027
−37.233
−19.393
1.00
24.28
O


ATOM
9017
ND2
ASN
B
60
−81.515
−38.172
−18.017
1.00
24.36
N


ATOM
9020
C
ASN
B
60
−77.898
−35.427
−16.234
1.00
21.17
C


ATOM
9021
O
ASN
B
60
−77.700
−34.273
−16.614
1.00
20.85
O


ATOM
9023
N
VAL
B
61
−77.745
−35.805
−14.968
1.00
20.60
N


ATOM
9024
CA
VAL
B
61
−77.382
−34.841
−13.942
1.00
20.48
C


ATOM
9026
CB
VAL
B
61
−77.277
−35.490
−12.551
1.00
20.26
C


ATOM
9028
CG1
VAL
B
61
−76.585
−34.562
−11.565
1.00
19.92
C


ATOM
9032
CG2
VAL
B
61
−78.668
−35.857
−12.049
1.00
20.66
C


ATOM
9036
C
VAL
B
61
−76.068
−34.208
−14.335
1.00
20.66
C


ATOM
9037
O
VAL
B
61
−75.871
−32.998
−14.182
1.00
20.74
O


ATOM
9039
N
GLN
B
62
−75.187
−35.048
−14.875
1.00
20.92
N


ATOM
9040
CA
GLN
B
62
−73.854
−34.636
−15.285
1.00
20.69
C


ATOM
9042
CB
GLN
B
62
−72.917
−35.847
−15.404
1.00
20.77
C


ATOM
9045
CG
GLN
B
62
−72.456
−36.369
−14.035
1.00
20.91
C


ATOM
9048
CD
GLN
B
62
−71.328
−37.383
−14.112
1.00
20.75
C


ATOM
9049
OE1
GLN
B
62
−70.512
−37.498
−13.192
1.00
20.93
O


ATOM
9050
NE2
GLN
B
62
−71.291
−38.135
−15.195
1.00
20.39
N


ATOM
9053
C
GLN
B
62
−73.900
−33.834
−16.567
1.00
20.32
C


ATOM
9054
O
GLN
B
62
−73.410
−32.712
−16.569
1.00
20.79
O


ATOM
9056
N
ARG
B
63
−74.512
−34.373
−17.625
1.00
19.87
N


ATOM
9057
CA
ARG
B
63
−74.520
−33.696
−18.940
1.00
19.81
C


ATOM
9059
CB
ARG
B
63
−75.240
−34.536
−20.013
1.00
19.75
C


ATOM
9062
CG
ARG
B
63
−74.492
−35.846
−20.345
1.00
21.69
C


ATOM
9065
CD
ARG
B
63
−75.158
−36.786
−21.368
1.00
24.89
C


ATOM
9068
NE
ARG
B
63
−74.674
−36.558
−22.741
1.00
29.32
N


ATOM
9070
CZ
ARG
B
63
−75.323
−35.878
−23.702
1.00
33.71
C


ATOM
9071
NH1
ARG
B
63
−76.537
−35.345
−23.510
1.00
36.67
N


ATOM
9074
NH2
ARG
B
63
−74.761
−35.736
−24.894
1.00
34.58
N


ATOM
9077
C
ARG
B
63
−75.099
−32.276
−18.822
1.00
19.39
C


ATOM
9078
O
ARG
B
63
−74.553
−31.310
−19.395
1.00
19.63
O


ATOM
9080
N
LEU
B
64
−76.155
−32.148
−18.017
1.00
18.62
N


ATOM
9081
CA
LEU
B
64
−76.811
−30.856
−17.753
1.00
17.96
C


ATOM
9083
CB
LEU
B
64
−78.136
−31.077
−17.008
1.00
17.97
C


ATOM
9086
CG
LEU
B
64
−79.264
−31.737
−17.809
1.00
17.08
C


ATOM
9088
CD1
LEU
B
64
−80.276
−32.354
−16.879
1.00
15.15
C


ATOM
9092
CD2
LEU
B
64
−79.904
−30.734
−18.737
1.00
15.30
C


ATOM
9096
C
LEU
B
64
−75.975
−29.847
−16.966
1.00
17.39
C


ATOM
9097
O
LEU
B
64
−76.370
−28.685
−16.825
1.00
17.38
O


ATOM
9099
N
GLY
B
65
−74.848
−30.286
−16.432
1.00
16.73
N


ATOM
9100
CA
GLY
B
65
−73.917
−29.374
−15.818
1.00
16.65
C


ATOM
9103
C
GLY
B
65
−74.077
−29.249
−14.319
1.00
16.68
C


ATOM
9104
O
GLY
B
65
−73.565
−28.302
−13.718
1.00
16.79
O


ATOM
9106
N
LEU
B
66
−74.758
−30.209
−13.702
1.00
16.60
N


ATOM
9107
CA
LEU
B
66
−74.978
−30.177
−12.264
1.00
16.58
C


ATOM
9109
CB
LEU
B
66
−76.465
−30.389
−11.957
1.00
16.35
C


ATOM
9112
CG
LEU
B
66
−77.363
−29.187
−12.234
1.00
15.57
C


ATOM
9114
CD1
LEU
B
66
−78.828
−29.602
−12.163
1.00
15.26
C


ATOM
9118
CD2
LEU
B
66
−77.063
−28.051
−11.270
1.00
13.66
C


ATOM
9122
C
LEU
B
66
−74.129
−31.203
−11.513
1.00
16.94
C


ATOM
9123
O
LEU
B
66
−74.152
−31.251
−10.279
1.00
17.50
O


ATOM
9125
N
GLY
B
67
−73.373
−32.017
−12.236
1.00
17.03
N


ATOM
9126
CA
GLY
B
67
−72.541
−33.036
−11.602
1.00
17.13
C


ATOM
9129
C
GLY
B
67
−71.642
−32.557
−10.461
1.00
17.08
C


ATOM
9130
O
GLY
B
67
−71.378
−33.317
−9.522
1.00
17.33
O


ATOM
9132
N
TYR
B
68
−71.159
−31.316
−10.541
1.00
16.83
N


ATOM
9133
CA
TYR
B
68
−70.217
−30.804
−9.552
1.00
16.64
C


ATOM
9135
CB
TYR
B
68
−69.654
−29.437
−9.951
1.00
16.25
C


ATOM
9138
CG
TYR
B
68
−70.609
−28.273
−9.802
1.00
13.51
C


ATOM
9139
CD1
TYR
B
68
−70.521
−27.411
−8.738
1.00
10.49
C


ATOM
9141
CE1
TYR
B
68
−71.407
−26.342
−8.608
1.00
10.25
C


ATOM
9143
CZ
TYR
B
68
−72.382
−26.135
−9.555
1.00
9.74
C


ATOM
9144
OH
TYR
B
68
−73.253
−25.086
−9.450
1.00
7.62
O


ATOM
9146
CE2
TYR
B
68
−72.484
−26.978
−10.625
1.00
11.14
C


ATOM
9148
CD2
TYR
B
68
−71.603
−28.037
−10.748
1.00
12.56
C


ATOM
9150
C
TYR
B
68
−70.828
−30.700
−8.172
1.00
17.79
C


ATOM
9151
O
TYR
B
68
−70.107
−30.811
−7.182
1.00
18.00
O


ATOM
9153
N
ARG
B
69
−72.146
−30.485
−8.090
1.00
18.85
N


ATOM
9154
CA
ARG
B
69
−72.784
−30.269
−6.789
1.00
19.46
C


ATOM
9156
CB
ARG
B
69
−73.708
−29.047
−6.819
1.00
19.23
C


ATOM
9159
CG
ARG
B
69
−75.030
−29.219
−7.509
1.00
18.83
C


ATOM
9162
CD
ARG
B
69
−76.053
−28.192
−6.985
1.00
17.73
C


ATOM
9165
NE
ARG
B
69
−75.642
−26.830
−7.297
1.00
16.01
N


ATOM
9167
CZ
ARG
B
69
−75.330
−25.889
−6.417
1.00
14.68
C


ATOM
9168
NH1
ARG
B
69
−75.400
−26.093
−5.112
1.00
14.99
N


ATOM
9171
NH2
ARG
B
69
−74.959
−24.707
−6.861
1.00
15.04
N


ATOM
9174
C
ARG
B
69
−73.490
−31.489
−6.234
1.00
20.50
C


ATOM
9175
O
ARG
B
69
−74.084
−31.418
−5.163
1.00
20.67
O


ATOM
9177
N
PHE
B
70
−73.416
−32.607
−6.959
1.00
22.04
N


ATOM
9178
CA
PHE
B
70
−73.853
−33.917
−6.450
1.00
23.03
C


ATOM
9180
CB
PHE
B
70
−75.081
−34.428
−7.211
1.00
22.94
C


ATOM
9183
CG
PHE
B
70
−76.236
−33.503
−7.157
1.00
21.78
C


ATOM
9184
CD1
PHE
B
70
−77.024
−33.441
−6.028
1.00
20.54
C


ATOM
9186
CE1
PHE
B
70
−78.076
−32.571
−5.959
1.00
20.32
C


ATOM
9188
CZ
PHE
B
70
−78.353
−31.752
−7.025
1.00
20.73
C


ATOM
9190
CE2
PHE
B
70
−77.564
−31.796
−8.160
1.00
21.24
C


ATOM
9192
CD2
PHE
B
70
−76.511
−32.665
−8.220
1.00
21.22
C


ATOM
9194
C
PHE
B
70
−72.765
−34.962
−6.570
1.00
24.33
C


ATOM
9195
O
PHE
B
70
−73.069
−36.142
−6.614
1.00
24.56
O


ATOM
9197
N
GLU
B
71
−71.500
−34.547
−6.624
1.00
26.01
N


ATOM
9198
CA
GLU
B
71
−70.413
−35.506
−6.804
1.00
27.04
C


ATOM
9200
CB
GLU
B
71
−69.042
−34.824
−6.885
1.00
27.48
C


ATOM
9203
CG
GLU
B
71
−67.851
−35.816
−6.794
1.00
28.94
C


ATOM
9206
CD
GLU
B
71
−66.491
−35.192
−7.081
1.00
30.38
C


ATOM
9207
OE1
GLU
B
71
−66.398
−34.192
−7.835
1.00
30.52
O


ATOM
9208
OE2
GLU
B
71
−65.501
−35.730
−6.544
1.00
32.16
O


ATOM
9209
C
GLU
B
71
−70.436
−36.531
−5.680
1.00
27.46
C


ATOM
9210
O
GLU
B
71
−70.449
−37.723
−5.937
1.00
27.67
O


ATOM
9212
N
SER
B
72
−70.464
−36.087
−4.435
1.00
28.04
N


ATOM
9213
CA
SER
B
72
−70.381
−37.044
−3.348
1.00
28.86
C


ATOM
9215
CB
SER
B
72
−70.388
−36.340
−1.983
1.00
29.09
C


ATOM
9218
OG
SER
B
72
−71.649
−35.751
−1.687
1.00
30.21
O


ATOM
9220
C
SER
B
72
−71.519
−38.058
−3.481
1.00
29.30
C


ATOM
9221
O
SER
B
72
−71.304
−39.265
−3.384
1.00
29.24
O


ATOM
9223
N
ASP
B
73
−72.719
−37.560
−3.756
1.00
30.03
N


ATOM
9224
CA
ASP
B
73
−73.897
−38.417
−3.874
1.00
30.63
C


ATOM
9226
CB
ASP
B
73
−75.184
−37.598
−4.116
1.00
30.74
C


ATOM
9229
CG
ASP
B
73
−75.419
−36.509
−3.052
1.00
31.84
C


ATOM
9230
OD1
ASP
B
73
−75.562
−36.830
−1.841
1.00
31.70
O


ATOM
9231
OD2
ASP
B
73
−75.473
−35.317
−3.443
1.00
33.81
O


ATOM
9232
C
ASP
B
73
−73.722
−39.443
−4.994
1.00
30.81
C


ATOM
9233
O
ASP
B
73
−74.130
−40.590
−4.835
1.00
31.39
O


ATOM
9235
N
ILE
B
74
−73.122
−39.031
−6.112
1.00
30.96
N


ATOM
9236
CA
ILE
B
74
−72.917
−39.912
−7.282
1.00
31.04
C


ATOM
9238
CB
ILE
B
74
−72.461
−39.114
−8.524
1.00
30.88
C


ATOM
9240
CG1
ILE
B
74
−73.585
−38.213
−9.018
1.00
30.99
C


ATOM
9243
CD1
ILE
B
74
−73.105
−37.120
−9.923
1.00
31.54
C


ATOM
9247
CG2
ILE
B
74
−72.052
−40.035
−9.642
1.00
29.85
C


ATOM
9251
C
ILE
B
74
−71.885
−41.005
−6.999
1.00
31.45
C


ATOM
9252
O
ILE
B
74
−72.098
−42.165
−7.347
1.00
31.31
O


ATOM
9254
N
ARG
B
75
−70.768
−40.628
−6.380
1.00
31.93
N


ATOM
9255
CA
ARG
B
75
−69.778
−41.601
−5.947
1.00
32.38
C


ATOM
9257
CB
ARG
B
75
−68.629
−40.959
−5.137
1.00
32.87
C


ATOM
9260
CG
ARG
B
75
−67.310
−40.751
−5.915
1.00
35.01
C


ATOM
9263
CD
ARG
B
75
−66.089
−40.489
−5.003
1.00
37.62
C


ATOM
9266
NE
ARG
B
75
−65.260
−41.687
−4.783
1.00
41.03
N


ATOM
9268
CZ
ARG
B
75
−64.461
−42.253
−5.700
1.00
44.29
C


ATOM
9269
NH1
ARG
B
75
−64.385
−41.751
−6.931
1.00
46.22
N


ATOM
9272
NH2
ARG
B
75
−63.741
−43.341
−5.404
1.00
44.30
N


ATOM
9275
C
ARG
B
75
−70.478
−42.673
−5.126
1.00
32.18
C


ATOM
9276
O
ARG
B
75
−70.307
−43.849
−5.398
1.00
32.19
O


ATOM
9278
N
ARG
B
76
−71.280
−42.275
−4.143
1.00
32.21
N


ATOM
9279
CA
ARG
B
76
−71.975
−43.256
−3.307
1.00
32.61
C


ATOM
9281
CB
ARG
B
76
−72.737
−42.582
−2.162
1.00
32.75
C


ATOM
9284
CG
ARG
B
76
−71.880
−42.320
−.929
1.00
33.44
C


ATOM
9287
CD
ARG
B
76
−72.720
−42.046
.334
1.00
34.16
C


ATOM
9290
NE
ARG
B
76
−73.797
−41.074
.124
1.00
34.53
N


ATOM
9292
CZ
ARG
B
76
−73.627
−39.765
−.063
1.00
34.38
C


ATOM
9293
NH1
ARG
B
76
−72.414
−39.219
−.092
1.00
34.34
N


ATOM
9296
NH2
ARG
B
76
−74.689
−38.993
−.241
1.00
34.71
N


ATOM
9299
C
ARG
B
76
−72.918
−44.159
−4.112
1.00
32.81
C


ATOM
9300
O
ARG
B
76
−72.863
−45.379
−3.995
1.00
32.86
O


ATOM
9302
N
ALA
B
77
−73.780
−43.559
−4.923
1.00
33.10
N


ATOM
9303
CA
ALA
B
77
−74.655
−44.307
−5.813
1.00
33.27
C


ATOM
9305
CB
ALA
B
77
−75.321
−43.366
−6.770
1.00
33.26
C


ATOM
9309
C
ALA
B
77
−73.884
−45.370
−6.578
1.00
33.80
C


ATOM
9310
O
ALA
B
77
−74.235
−46.542
−6.559
1.00
33.75
O


ATOM
9312
N
LEU
B
78
−72.816
−44.952
−7.242
1.00
34.77
N


ATOM
9313
CA
LEU
B
78
−71.987
−45.868
−8.019
1.00
35.41
C


ATOM
9315
CB
LEU
B
78
−70.845
−45.109
−8.702
1.00
35.01
C


ATOM
9318
CG
LEU
B
78
−71.220
−44.092
−9.782
1.00
34.37
C


ATOM
9320
CD1
LEU
B
78
−69.945
−43.502
−10.381
1.00
33.93
C


ATOM
9324
CD2
LEU
B
78
−72.109
−44.692
−10.873
1.00
32.94
C


ATOM
9328
C
LEU
B
78
−71.409
−46.974
−7.142
1.00
36.50
C


ATOM
9329
O
LEU
B
78
−71.336
−48.124
−7.552
1.00
36.87
O


ATOM
9331
N
ASP
B
79
−71.005
−46.619
−5.931
1.00
37.74
N


ATOM
9332
CA
ASP
B
79
−70.308
−47.545
−5.056
1.00
38.74
C


ATOM
9334
CB
ASP
B
79
−69.787
−46.810
−3.819
1.00
38.99
C


ATOM
9337
CG
ASP
B
79
−68.499
−47.386
−3.313
1.00
40.08
C


ATOM
9338
OD1
ASP
B
79
−68.494
−48.591
−2.976
1.00
42.32
O


ATOM
9339
OD2
ASP
B
79
−67.494
−46.638
−3.261
1.00
41.24
O


ATOM
9340
C
ASP
B
79
−71.207
−48.705
−4.645
1.00
39.42
C


ATOM
9341
O
ASP
B
79
−70.736
−49.835
−4.505
1.00
39.58
O


ATOM
9343
N
ARG
B
80
−72.495
−48.424
−4.450
1.00
40.26
N


ATOM
9344
CA
ARG
B
80
−73.471
−49.470
−4.143
1.00
40.92
C


ATOM
9346
CB
ARG
B
80
−74.823
−48.881
−3.716
1.00
41.46
C


ATOM
9349
CG
ARG
B
80
−74.871
−48.247
−2.308
1.00
42.87
C


ATOM
9352
CD
ARG
B
80
−76.316
−47.826
−1.936
1.00
44.62
C


ATOM
9355
NE
ARG
B
80
−76.946
−46.968
−2.949
1.00
45.87
N


ATOM
9357
CZ
ARG
B
80
−76.741
−45.651
−3.089
1.00
46.82
C


ATOM
9358
NH1
ARG
B
80
−75.905
−44.989
−2.282
1.00
46.27
N


ATOM
9361
NH2
ARG
B
80
−77.378
−44.988
−4.059
1.00
47.22
N


ATOM
9364
C
ARG
B
80
−73.667
−50.341
−5.371
1.00
40.77
C


ATOM
9365
O
ARG
B
80
−73.642
−51.563
−5.275
1.00
41.06
O


ATOM
9367
N
PHE
B
81
−73.863
−49.705
−6.520
1.00
40.56
N


ATOM
9368
CA
PHE
B
81
−73.986
−50.418
−7.787
1.00
40.59
C


ATOM
9370
CB
PHE
B
81
−73.984
−49.426
−8.956
1.00
40.73
C


ATOM
9373
CG
PHE
B
81
−73.898
−50.063
−10.323
1.00
40.84
C


ATOM
9374
CD1
PHE
B
81
−75.000
−50.668
−10.893
1.00
41.39
C


ATOM
9376
CE1
PHE
B
81
−74.925
−51.236
−12.167
1.00
41.67
C


ATOM
9378
CZ
PHE
B
81
−73.741
−51.191
−12.877
1.00
41.33
C


ATOM
9380
CE2
PHE
B
81
−72.638
−50.580
−12.326
1.00
41.19
C


ATOM
9382
CD2
PHE
B
81
−72.720
−50.012
−11.057
1.00
41.39
C


ATOM
9384
C
PHE
B
81
−72.871
−51.441
−7.953
1.00
40.47
C


ATOM
9385
O
PHE
B
81
−73.134
−52.583
−8.311
1.00
40.85
O


ATOM
9387
N
VAL
B
82
−71.633
−51.051
−7.683
1.00
40.26
N


ATOM
9388
CA
VAL
B
82
−70.522
−51.986
−7.821
1.00
40.24
C


ATOM
9390
CB
VAL
B
82
−69.173
−51.386
−7.379
1.00
40.28
C


ATOM
9392
CG1
VAL
B
82
−68.164
−52.493
−7.063
1.00
39.98
C


ATOM
9396
CG2
VAL
B
82
−68.644
−50.462
−8.451
1.00
40.09
C


ATOM
9400
C
VAL
B
82
−70.788
−53.230
−7.005
1.00
40.16
C


ATOM
9401
O
VAL
B
82
−70.880
−54.313
−7.559
1.00
40.13
O


ATOM
9403
N
SER
B
83
−70.953
−53.056
−5.698
1.00
40.26
N


ATOM
9404
CA
SER
B
83
−71.070
−54.174
−4.749
1.00
40.43
C


ATOM
9406
CB
SER
B
83
−70.797
−53.666
−3.331
1.00
40.54
C


ATOM
9409
OG
SER
B
83
−71.256
−52.330
−3.191
1.00
40.89
O


ATOM
9411
C
SER
B
83
−72.415
−54.912
−4.831
1.00
40.34
C


ATOM
9412
O
SER
B
83
−73.137
−55.046
−3.845
1.00
39.99
O


ATOM
9414
N
SER
B
84
−72.698
−55.400
−6.038
1.00
40.57
N


ATOM
9415
CA
SER
B
84
−73.902
−56.154
−6.411
1.00
40.63
C


ATOM
9417
CB
SER
B
84
−75.154
−55.679
−5.651
1.00
40.56
C


ATOM
9420
OG
SER
B
84
−75.292
−54.268
−5.663
1.00
39.82
O


ATOM
9422
C
SER
B
84
−74.104
−56.026
−7.940
1.00
40.85
C


ATOM
9423
O
SER
B
84
−75.104
−55.462
−8.395
1.00
41.24
O


ATOM
9425
N
GLY
B
85
−73.136
−56.524
−8.720
1.00
40.75
N


ATOM
9426
CA
GLY
B
85
−73.201
−56.506
−10.191
1.00
40.64
C


ATOM
9429
C
GLY
B
85
−73.225
−55.120
−10.815
1.00
40.64
C


ATOM
9430
O
GLY
B
85
−74.133
−54.794
−11.590
1.00
40.32
O


ATOM
9432
N
THR
B
93
−74.847
−57.360
−18.759
1.00
36.30
N


ATOM
9433
CA
THR
B
93
−75.593
−58.129
−19.774
1.00
35.99
C


ATOM
9435
CB
THR
B
93
−76.251
−59.379
−19.152
1.00
35.87
C


ATOM
9437
OG1
THR
B
93
−76.347
−60.389
−20.158
1.00
35.64
O


ATOM
9439
CG2
THR
B
93
−77.646
−59.062
−18.548
1.00
34.95
C


ATOM
9443
C
THR
B
93
−76.625
−57.274
−20.583
1.00
35.90
C


ATOM
9444
O
THR
B
93
−77.764
−57.688
−20.843
1.00
35.74
O


ATOM
9446
N
SER
B
94
−76.175
−56.076
−20.963
1.00
35.58
N


ATOM
9447
CA
SER
B
94
−76.874
−55.150
−21.866
1.00
34.97
C


ATOM
9449
CB
SER
B
94
−78.074
−54.472
−21.190
1.00
35.06
C


ATOM
9452
OG
SER
B
94
−77.702
−53.260
−20.541
1.00
34.37
O


ATOM
9454
C
SER
B
94
−75.831
−54.097
−22.232
1.00
34.39
C


ATOM
9455
O
SER
B
94
−75.212
−53.518
−21.341
1.00
34.11
O


ATOM
9457
N
LEU
B
95
−75.619
−53.864
−23.522
1.00
33.77
N


ATOM
9458
CA
LEU
B
95
−74.490
−53.045
−23.952
1.00
33.33
C


ATOM
9460
CB
LEU
B
95
−74.373
−53.010
−25.475
1.00
33.30
C


ATOM
9463
CG
LEU
B
95
−73.154
−52.246
−26.006
1.00
33.27
C


ATOM
9465
CD1
LEU
B
95
−71.909
−52.455
−25.137
1.00
32.89
C


ATOM
9469
CD2
LEU
B
95
−72.864
−52.649
−27.438
1.00
33.19
C


ATOM
9473
C
LEU
B
95
−74.551
−51.623
−23.413
1.00
33.00
C


ATOM
9474
O
LEU
B
95
−73.588
−51.151
−22.809
1.00
32.83
O


ATOM
9476
N
HIS
B
96
−75.678
−50.949
−23.633
1.00
32.64
N


ATOM
9477
CA
HIS
B
96
−75.853
−49.576
−23.171
1.00
32.39
C


ATOM
9479
CB
HIS
B
96
−77.246
−49.069
−23.527
1.00
32.64
C


ATOM
9482
CG
HIS
B
96
−77.528
−47.689
−23.025
1.00
34.16
C


ATOM
9483
ND1
HIS
B
96
−76.565
−46.703
−22.981
1.00
36.21
N


ATOM
9485
CE1
HIS
B
96
−77.096
−45.594
−22.496
1.00
36.92
C


ATOM
9487
NE2
HIS
B
96
−78.371
−45.827
−22.227
1.00
37.06
N


ATOM
9489
CD2
HIS
B
96
−78.664
−47.130
−22.548
1.00
35.43
C


ATOM
9491
C
HIS
B
96
−75.612
−49.445
−21.665
1.00
31.78
C


ATOM
9492
O
HIS
B
96
−74.936
−48.523
−21.215
1.00
31.95
O


ATOM
9494
N
GLY
B
97
−76.156
−50.372
−20.887
1.00
31.10
N


ATOM
9495
CA
GLY
B
97
−75.907
−50.396
−19.446
1.00
30.52
C


ATOM
9498
C
GLY
B
97
−74.434
−50.532
−19.085
1.00
29.95
C


ATOM
9499
O
GLY
B
97
−73.936
−49.797
−18.241
1.00
30.21
O


ATOM
9501
N
THR
B
98
−73.740
−51.470
−19.726
1.00
29.15
N


ATOM
9502
CA
THR
B
98
−72.314
−51.690
−19.490
1.00
28.55
C


ATOM
9504
CB
THR
B
98
−71.803
−52.955
−20.248
1.00
28.48
C


ATOM
9506
OG1
THR
B
98
−72.678
−54.066
−19.997
1.00
28.41
O


ATOM
9508
CG2
THR
B
98
−70.405
−53.331
−19.816
1.00
27.74
C


ATOM
9512
C
THR
B
98
−71.492
−50.452
−19.894
1.00
28.31
C


ATOM
9513
O
THR
B
98
−70.658
−49.979
−19.127
1.00
28.13
O


ATOM
9515
N
ALA
B
99
−71.750
−49.910
−21.080
1.00
28.04
N


ATOM
9516
CA
ALA
B
99
−70.984
−48.758
−21.578
1.00
27.88
C


ATOM
9518
CB
ALA
B
99
−71.387
−48.406
−23.014
1.00
27.78
C


ATOM
9522
C
ALA
B
99
−71.127
−47.538
−20.684
1.00
27.62
C


ATOM
9523
O
ALA
B
99
−70.135
−46.885
−20.361
1.00
27.84
O


ATOM
9525
N
LEU
B
100
−72.357
−47.233
−20.290
1.00
27.20
N


ATOM
9526
CA
LEU
B
100
−72.613
−46.088
−19.422
1.00
27.08
C


ATOM
9528
CB
LEU
B
100
−74.122
−45.900
−19.230
1.00
26.95
C


ATOM
9531
CG
LEU
B
100
−74.560
−44.715
−18.367
1.00
26.27
C


ATOM
9533
CD1
LEU
B
100
−73.753
−43.478
−18.734
1.00
26.19
C


ATOM
9537
CD2
LEU
B
100
−76.052
−44.445
−18.504
1.00
25.20
C


ATOM
9541
C
LEU
B
100
−71.928
−46.221
−18.046
1.00
27.23
C


ATOM
9542
O
LEU
B
100
−71.368
−45.243
−17.523
1.00
27.16
O


ATOM
9544
N
SER
B
101
−71.980
−47.429
−17.472
1.00
27.00
N


ATOM
9545
CA
SER
B
101
−71.475
−47.682
−16.117
1.00
26.63
C


ATOM
9547
CB
SER
B
101
−72.039
−48.992
−15.568
1.00
26.49
C


ATOM
9550
OG
SER
B
101
−71.758
−50.063
−16.442
1.00
26.38
O


ATOM
9552
C
SER
B
101
−69.956
−47.740
−16.090
1.00
26.47
C


ATOM
9553
O
SER
B
101
−69.326
−47.324
−15.121
1.00
26.35
O


ATOM
9555
N
PHE
B
102
−69.381
−48.288
−17.154
1.00
26.29
N


ATOM
9556
CA
PHE
B
102
−67.934
−48.332
−17.322
1.00
25.91
C


ATOM
9558
CB
PHE
B
102
−67.588
−49.005
−18.652
1.00
25.96
C


ATOM
9561
CG
PHE
B
102
−66.133
−49.020
−18.958
1.00
25.81
C


ATOM
9562
CD1
PHE
B
102
−65.360
−50.115
−18.633
1.00
25.96
C


ATOM
9564
CE1
PHE
B
102
−63.999
−50.122
−18.909
1.00
26.94
C


ATOM
9566
CZ
PHE
B
102
−63.403
−49.026
−19.519
1.00
26.27
C


ATOM
9568
CE2
PHE
B
102
−64.174
−47.930
−19.848
1.00
26.19
C


ATOM
9570
CD2
PHE
B
102
−65.531
−47.933
−19.573
1.00
26.04
C


ATOM
9572
C
PHE
B
102
−67.416
−46.911
−17.315
1.00
25.56
C


ATOM
9573
O
PHE
B
102
−66.486
−46.564
−16.568
1.00
25.37
O


ATOM
9575
N
ARG
B
103
−68.050
−46.095
−18.152
1.00
25.20
N


ATOM
9576
CA
ARG
B
103
−67.663
−44.696
−18.322
1.00
25.09
C


ATOM
9578
CB
ARG
B
103
−68.510
−44.026
−19.400
1.00
25.12
C


ATOM
9581
CG
ARG
B
103
−68.194
−42.561
−19.572
1.00
25.41
C


ATOM
9584
CD
ARG
B
103
−68.744
−42.035
−20.889
1.00
26.68
C


ATOM
9587
NE
ARG
B
103
−70.197
−41.843
−20.877
1.00
27.49
N


ATOM
9589
CZ
ARG
B
103
−70.826
−40.851
−20.246
1.00
26.56
C


ATOM
9590
NH1
ARG
B
103
−70.139
−39.963
−19.531
1.00
27.11
N


ATOM
9593
NH2
ARG
B
103
−72.148
−40.759
−20.310
1.00
25.20
N


ATOM
9596
C
ARG
B
103
−67.785
−43.898
−17.033
1.00
24.61
C


ATOM
9597
O
ARG
B
103
−66.865
−43.168
−16.659
1.00
24.67
O


ATOM
9599
N
LEU
B
104
−68.927
−44.013
−16.371
1.00
23.79
N


ATOM
9600
CA
LEU
B
104
−69.125
−43.263
−15.148
1.00
23.23
C


ATOM
9602
CB
LEU
B
104
−70.591
−43.342
−14.693
1.00
23.01
C


ATOM
9605
CG
LEU
B
104
−71.607
−42.620
−15.584
1.00
21.36
C


ATOM
9607
CD1
LEU
B
104
−73.002
−42.860
−15.067
1.00
19.31
C


ATOM
9611
CD2
LEU
B
104
−71.310
−41.157
−15.633
1.00
19.57
C


ATOM
9615
C
LEU
B
104
−68.156
−43.781
−14.071
1.00
23.15
C


ATOM
9616
O
LEU
B
104
−67.445
−43.007
−13.423
1.00
23.40
O


ATOM
9618
N
LEU
B
105
−68.106
−45.091
−13.894
1.00
22.79
N


ATOM
9619
CA
LEU
B
105
−67.203
−45.659
−12.916
1.00
22.40
C


ATOM
9621
CB
LEU
B
105
−67.303
−47.178
−12.912
1.00
22.42
C


ATOM
9624
CG
LEU
B
105
−68.505
−47.709
−12.163
1.00
21.82
C


ATOM
9626
CD1
LEU
B
105
−68.810
−49.120
−12.591
1.00
21.95
C


ATOM
9630
CD2
LEU
B
105
−68.201
−47.648
−10.692
1.00
22.14
C


ATOM
9634
C
LEU
B
105
−65.767
−45.239
−13.196
1.00
22.33
C


ATOM
9635
O
LEU
B
105
−65.049
−44.823
−12.275
1.00
22.13
O


ATOM
9637
N
ARG
B
106
−65.331
−45.342
−14.451
1.00
22.08
N


ATOM
9638
CA
ARG
B
106
−63.953
−44.973
−14.730
1.00
22.20
C


ATOM
9640
CB
ARG
B
106
−63.521
−45.266
−16.151
1.00
22.33
C


ATOM
9643
CG
ARG
B
106
−62.075
−44.827
−16.329
1.00
23.60
C


ATOM
9646
CD
ARG
B
106
−61.383
−45.445
−17.513
1.00
24.71
C


ATOM
9649
NE
ARG
B
106
−61.078
−46.852
−17.319
1.00
24.66
N


ATOM
9651
CZ
ARG
B
106
−60.418
−47.582
−18.206
1.00
25.75
C


ATOM
9652
NH1
ARG
B
106
−59.995
−47.028
−19.337
1.00
26.41
N


ATOM
9655
NH2
ARG
B
106
−60.182
−48.866
−17.971
1.00
26.54
N


ATOM
9658
C
ARG
B
106
−63.732
−43.504
−14.438
1.00
21.75
C


ATOM
9659
O
ARG
B
106
−62.801
−43.145
−13.725
1.00
21.50
O


ATOM
9661
N
GLN
B
107
−64.609
−42.680
−15.003
1.00
21.69
N


ATOM
9662
CA
GLN
B
107
−64.630
−41.239
−14.794
1.00
21.64
C


ATOM
9664
CB
GLN
B
107
−65.964
−40.664
−15.261
1.00
21.71
C


ATOM
9667
CG
GLN
B
107
−66.169
−39.178
−14.929
1.00
21.97
C


ATOM
9670
CD
GLN
B
107
−67.588
−38.723
−15.172
1.00
21.54
C


ATOM
9671
OE1
GLN
B
107
−68.355
−39.365
−15.906
1.00
20.08
O


ATOM
9672
NE2
GLN
B
107
−67.948
−37.600
−14.556
1.00
21.45
N


ATOM
9675
C
GLN
B
107
−64.454
−40.850
−13.352
1.00
21.70
C


ATOM
9676
O
GLN
B
107
−63.776
−39.865
−13.078
1.00
22.14
O


ATOM
9678
N
HIS
B
108
−65.091
−41.599
−12.450
1.00
21.53
N


ATOM
9679
CA
HIS
B
108
−65.049
−41.326
−11.019
1.00
21.63
C


ATOM
9681
CB
HIS
B
108
−66.447
−41.535
−10.429
1.00
21.47
C


ATOM
9684
CG
HIS
B
108
−67.416
−40.445
−10.752
1.00
20.91
C


ATOM
9685
ND1
HIS
B
108
−67.541
−39.315
−9.976
1.00
20.66
N


ATOM
9687
CE1
HIS
B
108
−68.476
−38.535
−10.490
1.00
20.90
C


ATOM
9689
NE2
HIS
B
108
−68.966
−39.118
−11.567
1.00
19.67
N


ATOM
9691
CD2
HIS
B
108
−68.326
−40.320
−11.747
1.00
20.76
C


ATOM
9693
C
HIS
B
108
−64.024
−42.209
−10.269
1.00
22.22
C


ATOM
9694
O
HIS
B
108
−64.220
−42.553
−9.104
1.00
22.17
O


ATOM
9696
N
GLY
B
109
−62.950
−42.613
−10.933
1.00
22.91
N


ATOM
9697
CA
GLY
B
109
−61.846
−43.288
−10.245
1.00
23.85
C


ATOM
9700
C
GLY
B
109
−61.986
−44.748
−9.800
1.00
24.50
C


ATOM
9701
O
GLY
B
109
−61.053
−45.306
−9.189
1.00
24.12
O


ATOM
9703
N
PHE
B
110
−63.128
−45.371
−10.094
1.00
25.16
N


ATOM
9704
CA
PHE
B
110
−63.309
−46.791
−9.800
1.00
25.67
C


ATOM
9706
CB
PHE
B
110
−64.782
−47.187
−9.891
1.00
25.80
C


ATOM
9709
CG
PHE
B
110
−65.625
−46.656
−8.772
1.00
26.12
C


ATOM
9710
CD1
PHE
B
110
−65.564
−47.230
−7.510
1.00
27.13
C


ATOM
9712
CE1
PHE
B
110
−66.353
−46.751
−6.468
1.00
27.42
C


ATOM
9714
CZ
PHE
B
110
−67.215
−45.691
−6.692
1.00
26.93
C


ATOM
9716
CE2
PHE
B
110
−67.284
−45.119
−7.954
1.00
26.49
C


ATOM
9718
CD2
PHE
B
110
−66.494
−45.601
−8.981
1.00
25.96
C


ATOM
9720
C
PHE
B
110
−62.505
−47.615
−10.793
1.00
25.92
C


ATOM
9721
O
PHE
B
110
−62.232
−47.157
−11.898
1.00
26.41
O


ATOM
9723
N
GLU
B
111
−62.134
−48.832
−10.406
1.00
26.06
N


ATOM
9724
CA
GLU
B
111
−61.403
−49.724
−11.304
1.00
26.17
C


ATOM
9726
CB
GLU
B
111
−60.444
−50.609
−10.511
1.00
26.50
C


ATOM
9729
CG
GLU
B
111
−59.372
−51.272
−11.374
1.00
28.11
C


ATOM
9732
CD
GLU
B
111
−58.607
−52.377
−10.646
1.00
30.70
C


ATOM
9733
OE1
GLU
B
111
−58.660
−52.436
−9.390
1.00
31.99
O


ATOM
9734
OE2
GLU
B
111
−57.948
−53.190
−11.338
1.00
32.09
O


ATOM
9735
C
GLU
B
111
−62.355
−50.598
−12.130
1.00
25.60
C


ATOM
9736
O
GLU
B
111
−63.116
−51.388
−11.585
1.00
25.48
O


ATOM
9738
N
VAL
B
112
−62.314
−50.439
−13.447
1.00
25.24
N


ATOM
9739
CA
VAL
B
112
−63.026
−51.331
−14.357
1.00
24.90
C


ATOM
9741
CB
VAL
B
112
−64.308
−50.712
−14.908
1.00
24.92
C


ATOM
9743
CG1
VAL
B
112
−65.268
−50.437
−13.771
1.00
25.26
C


ATOM
9747
CG2
VAL
B
112
−63.999
−49.450
−15.706
1.00
24.77
C


ATOM
9751
C
VAL
B
112
−62.144
−51.701
−15.522
1.00
24.66
C


ATOM
9752
O
VAL
B
112
−61.217
−50.964
−15.862
1.00
23.80
O


ATOM
9754
N
SER
B
113
−62.467
−52.842
−16.132
1.00
24.88
N


ATOM
9755
CA
SER
B
113
−61.604
−53.494
−17.109
1.00
25.21
C


ATOM
9757
CB
SER
B
113
−61.286
−54.913
−16.654
1.00
24.94
C


ATOM
9760
OG
SER
B
113
−60.357
−55.523
−17.528
1.00
24.31
O


ATOM
9762
C
SER
B
113
−62.233
−53.532
−18.492
1.00
25.88
C


ATOM
9763
O
SER
B
113
−63.446
−53.651
−18.630
1.00
25.86
O


ATOM
9765
N
GLN
B
114
−61.406
−53.438
−19.526
1.00
26.92
N


ATOM
9766
CA
GLN
B
114
−61.928
−53.491
−20.880
1.00
27.83
C


ATOM
9768
CB
GLN
B
114
−60.846
−53.194
−21.917
1.00
27.75
C


ATOM
9771
CG
GLN
B
114
−59.559
−53.952
−21.726
1.00
28.05
C


ATOM
9774
CD
GLN
B
114
−58.739
−54.077
−23.007
1.00
28.39
C


ATOM
9775
OE1
GLN
B
114
−59.086
−53.515
−24.051
1.00
29.05
O


ATOM
9776
NE2
GLN
B
114
−57.637
−54.808
−22.925
1.00
27.62
N


ATOM
9779
C
GLN
B
114
−62.615
−54.827
−21.158
1.00
28.78
C


ATOM
9780
O
GLN
B
114
−63.484
−54.905
−22.022
1.00
29.10
O


ATOM
9782
N
GLU
B
115
−62.248
−55.861
−20.401
1.00
29.99
N


ATOM
9783
CA
GLU
B
115
−62.888
−57.181
−20.505
1.00
30.89
C


ATOM
9785
CB
GLU
B
115
−62.252
−58.202
−19.549
1.00
31.10
C


ATOM
9788
CG
GLU
B
115
−60.740
−58.394
−19.697
1.00
32.35
C


ATOM
9791
CD
GLU
B
115
−60.320
−58.796
−21.106
1.00
34.20
C


ATOM
9792
OE1
GLU
B
115
−60.965
−59.699
−21.688
1.00
34.94
O


ATOM
9793
OE2
GLU
B
115
−59.348
−58.201
−21.634
1.00
35.60
O


ATOM
9794
C
GLU
B
115
−64.380
−57.120
−20.224
1.00
31.26
C


ATOM
9795
O
GLU
B
115
−65.125
−57.971
−20.691
1.00
31.54
O


ATOM
9797
N
ALA
B
116
−64.825
−56.127
−19.465
1.00
31.91
N


ATOM
9798
CA
ALA
B
116
−66.255
−55.937
−19.259
1.00
32.69
C


ATOM
9800
CB
ALA
B
116
−66.521
−54.648
−18.513
1.00
32.62
C


ATOM
9804
C
ALA
B
116
−66.999
−55.950
−20.597
1.00
33.33
C


ATOM
9805
O
ALA
B
116
−68.156
−56.364
−20.668
1.00
33.26
O


ATOM
9807
N
PHE
B
117
−66.316
−55.521
−21.655
1.00
34.16
N


ATOM
9808
CA
PHE
B
117
−66.889
−55.498
−22.996
1.00
34.98
C


ATOM
9810
CB
PHE
B
117
−66.310
−54.314
−23.766
1.00
35.02
C


ATOM
9813
CG
PHE
B
117
−66.868
−52.997
−23.345
1.00
35.06
C


ATOM
9814
CD1
PHE
B
117
−66.061
−52.046
−22.729
1.00
35.33
C


ATOM
9816
CE1
PHE
B
117
−66.576
−50.825
−22.342
1.00
35.16
C


ATOM
9818
CZ
PHE
B
117
−67.907
−50.544
−22.570
1.00
35.37
C


ATOM
9820
CE2
PHE
B
117
−68.723
−51.488
−23.187
1.00
35.28
C


ATOM
9822
CD2
PHE
B
117
−68.201
−52.701
−23.571
1.00
34.68
C


ATOM
9824
C
PHE
B
117
−66.676
−56.766
−23.830
1.00
35.86
C


ATOM
9825
O
PHE
B
117
−66.934
−56.755
−25.037
1.00
35.91
O


ATOM
9827
N
SER
B
118
−66.215
−57.853
−23.212
1.00
36.98
N


ATOM
9828
CA
SER
B
118
−65.924
−59.086
−23.960
1.00
37.76
C


ATOM
9830
CB
SER
B
118
−64.836
−59.907
−23.262
1.00
37.73
C


ATOM
9833
OG
SER
B
118
−65.272
−60.346
−21.987
1.00
37.77
O


ATOM
9835
C
SER
B
118
−67.178
−59.939
−24.215
1.00
38.48
C


ATOM
9836
O
SER
B
118
−67.207
−60.724
−25.157
1.00
38.42
O


ATOM
9838
N
GLY
B
119
−68.221
−59.765
−23.407
1.00
39.51
N


ATOM
9839
CA
GLY
B
119
−69.481
−60.473
−23.629
1.00
40.51
C


ATOM
9842
C
GLY
B
119
−70.287
−60.034
−24.848
1.00
41.51
C


ATOM
9843
O
GLY
B
119
−71.444
−60.433
−25.002
1.00
41.53
O


ATOM
9845
N
PHE
B
120
−69.688
−59.210
−25.709
1.00
42.79
N


ATOM
9846
CA
PHE
B
120
−70.382
−58.621
−26.864
1.00
43.81
C


ATOM
9848
CB
PHE
B
120
−70.643
−57.121
−26.601
1.00
43.76
C


ATOM
9851
CG
PHE
B
120
−71.367
−56.853
−25.293
1.00
43.65
C


ATOM
9852
CD1
PHE
B
120
−72.762
−56.874
−25.230
1.00
43.48
C


ATOM
9854
CE1
PHE
B
120
−73.437
−56.650
−24.023
1.00
43.03
C


ATOM
9856
CZ
PHE
B
120
−72.717
−56.413
−22.865
1.00
43.09
C


ATOM
9858
CE2
PHE
B
120
−71.323
−56.397
−22.909
1.00
43.38
C


ATOM
9860
CD2
PHE
B
120
−70.655
−56.619
−24.120
1.00
43.41
C


ATOM
9862
C
PHE
B
120
−69.607
−58.832
−28.175
1.00
44.81
C


ATOM
9863
O
PHE
B
120
−69.930
−58.238
−29.205
1.00
44.65
O


ATOM
9865
N
LYS
B
121
−68.602
−59.705
−28.126
1.00
46.19
N


ATOM
9866
CA
LYS
B
121
−67.757
−60.004
−29.272
1.00
47.31
C


ATOM
9868
CB
LYS
B
121
−66.275
−59.928
−28.870
1.00
47.47
C


ATOM
9871
CG
LYS
B
121
−65.743
−58.495
−28.637
1.00
48.13
C


ATOM
9874
CD
LYS
B
121
−64.532
−58.439
−27.675
1.00
48.89
C


ATOM
9877
CE
LYS
B
121
−63.261
−59.094
−28.241
1.00
49.20
C


ATOM
9880
NZ
LYS
B
121
−62.541
−58.232
−29.218
1.00
49.04
N


ATOM
9884
C
LYS
B
121
−68.096
−61.400
−29.810
1.00
48.15
C


ATOM
9885
O
LYS
B
121
−68.199
−62.361
−29.043
1.00
48.36
O


ATOM
9887
N
ASP
B
122
−68.270
−61.510
−31.126
1.00
49.04
N


ATOM
9888
CA
ASP
B
122
−68.611
−62.788
−31.754
1.00
49.60
C


ATOM
9890
CB
ASP
B
122
−69.070
−62.596
−33.217
1.00
49.52
C


ATOM
9893
CG
ASP
B
122
−67.989
−62.015
−34.126
1.00
49.18
C


ATOM
9894
OD1
ASP
B
122
−66.792
−62.306
−33.938
1.00
48.96
O


ATOM
9895
OD2
ASP
B
122
−68.351
−61.268
−35.057
1.00
48.82
O


ATOM
9896
C
ASP
B
122
−67.455
−63.785
−31.651
1.00
50.31
C


ATOM
9897
O
ASP
B
122
−66.369
−63.441
−31.171
1.00
50.33
O


ATOM
9899
N
GLN
B
123
−67.705
−65.014
−32.098
1.00
51.05
N


ATOM
9900
CA
GLN
B
123
−66.716
−66.095
−32.057
1.00
51.61
C


ATOM
9902
CB
GLN
B
123
−67.263
−67.330
−32.785
1.00
51.82
C


ATOM
9905
CG
GLN
B
123
−68.483
−67.986
−32.117
1.00
52.32
C


ATOM
9908
CD
GLN
B
123
−68.117
−69.137
−31.184
1.00
52.64
C


ATOM
9909
OE1
GLN
B
123
−67.146
−69.062
−30.432
1.00
53.02
O


ATOM
9910
NE2
GLN
B
123
−68.904
−70.207
−31.230
1.00
52.04
N


ATOM
9913
C
GLN
B
123
−65.358
−65.694
−32.659
1.00
51.78
C


ATOM
9914
O
GLN
B
123
−64.309
−66.000
−32.090
1.00
51.67
O


ATOM
9916
N
ASN
B
124
−65.388
−65.004
−33.799
1.00
52.05
N


ATOM
9917
CA
ASN
B
124
−64.166
−64.547
−34.473
1.00
52.25
C


ATOM
9919
CB
ASN
B
124
−64.486
−64.042
−35.884
1.00
52.25
C


ATOM
9922
CG
ASN
B
124
−64.911
−65.158
−36.819
1.00
51.88
C


ATOM
9923
OD1
ASN
B
124
−65.918
−65.825
−36.592
1.00
51.43
O


ATOM
9924
ND2
ASN
B
124
−64.144
−65.364
−37.880
1.00
51.23
N


ATOM
9927
C
ASN
B
124
−63.382
−63.466
−33.716
1.00
52.43
C


ATOM
9928
O
ASN
B
124
−62.189
−63.287
−33.959
1.00
52.33
O


ATOM
9930
N
GLY
B
125
−64.051
−62.747
−32.815
1.00
52.66
N


ATOM
9931
CA
GLY
B
125
−63.396
−61.745
−31.968
1.00
52.68
C


ATOM
9934
C
GLY
B
125
−63.916
−60.329
−32.135
1.00
52.63
C


ATOM
9935
O
GLY
B
125
−63.539
−59.447
−31.367
1.00
52.57
O


ATOM
9937
N
ASN
B
126
−64.782
−60.117
−33.129
1.00
52.54
N


ATOM
9938
CA
ASN
B
126
−65.343
−58.793
−33.443
1.00
52.41
C


ATOM
9940
CB
ASN
B
126
−65.575
−58.671
−34.949
1.00
52.40
C


ATOM
9943
CG
ASN
B
126
−64.322
−58.951
−35.751
1.00
52.64
C


ATOM
9944
OD1
ASN
B
126
−63.426
−59.665
−35.297
1.00
52.77
O


ATOM
9945
ND2
ASN
B
126
−64.249
−58.390
−36.952
1.00
53.00
N


ATOM
9948
C
ASN
B
126
−66.656
−58.524
−32.712
1.00
52.10
C


ATOM
9949
O
ASN
B
126
−67.253
−59.432
−32.153
1.00
52.27
O


ATOM
9951
N
PHE
B
127
−67.111
−57.279
−32.724
1.00
51.61
N


ATOM
9952
CA
PHE
B
127
−68.362
−56.936
−32.055
1.00
51.23
C


ATOM
9954
CB
PHE
B
127
−68.506
−55.416
−31.905
1.00
51.21
C


ATOM
9957
CG
PHE
B
127
−67.702
−54.848
−30.770
1.00
50.58
C


ATOM
9958
CD1
PHE
B
127
−66.513
−54.192
−31.003
1.00
50.15
C


ATOM
9960
CE1
PHE
B
127
−65.777
−53.686
−29.948
1.00
50.15
C


ATOM
9962
CZ
PHE
B
127
−66.225
−53.839
−28.647
1.00
49.71
C


ATOM
9964
CE2
PHE
B
127
−67.399
−54.496
−28.405
1.00
49.50
C


ATOM
9966
CD2
PHE
B
127
−68.132
−54.999
−29.459
1.00
49.99
C


ATOM
9968
C
PHE
B
127
−69.537
−57.505
−32.821
1.00
50.99
C


ATOM
9969
O
PHE
B
127
−69.468
−57.641
−34.040
1.00
50.92
O


ATOM
9971
N
LEU
B
128
−70.609
−57.840
−32.106
1.00
50.83
N


ATOM
9972
CA
LEU
B
128
−71.799
−58.422
−32.731
1.00
50.76
C


ATOM
9974
CB
LEU
B
128
−72.780
−58.953
−31.669
1.00
50.80
C


ATOM
9977
CG
LEU
B
128
−72.341
−60.098
−30.734
1.00
50.91
C


ATOM
9979
CD1
LEU
B
128
−73.483
−60.532
−29.813
1.00
50.61
C


ATOM
9983
CD2
LEU
B
128
−71.818
−61.303
−31.497
1.00
50.79
C


ATOM
9987
C
LEU
B
128
−72.500
−57.407
−33.646
1.00
50.55
C


ATOM
9988
O
LEU
B
128
−73.105
−56.455
−33.171
1.00
50.34
O


ATOM
9990
N
GLU
B
129
−72.402
−57.628
−34.957
1.00
50.50
N


ATOM
9991
CA
GLU
B
129
−73.038
−56.783
−35.982
1.00
50.48
C


ATOM
9993
CB
GLU
B
129
−73.150
−57.551
−37.310
1.00
50.68
C


ATOM
9996
CG
GLU
B
129
−72.262
−57.024
−38.434
1.00
51.36
C


ATOM
9999
CD
GLU
B
129
−72.799
−55.745
−39.060
1.00
51.88
C


ATOM
10000
OE1
GLU
B
129
−72.110
−54.707
−38.961
1.00
52.72
O


ATOM
10001
OE2
GLU
B
129
−73.906
−55.773
−39.643
1.00
51.58
O


ATOM
10002
C
GLU
B
129
−74.428
−56.264
−35.629
1.00
50.25
C


ATOM
10003
O
GLU
B
129
−74.737
−55.110
−35.899
1.00
50.18
O


ATOM
10005
N
ASN
B
130
−75.263
−57.125
−35.047
1.00
50.06
N


ATOM
10006
CA
ASN
B
130
−76.678
−56.806
−34.785
1.00
49.71
C


ATOM
10008
CB
ASN
B
130
−77.477
−58.100
−34.550
1.00
49.69
C


ATOM
10011
CG
ASN
B
130
−77.053
−58.837
−33.291
1.00
49.39
C


ATOM
10012
OD1
ASN
B
130
−76.357
−59.848
−33.361
1.00
48.74
O


ATOM
10013
ND2
ASN
B
130
−77.468
−58.329
−32.134
1.00
48.99
N


ATOM
10016
C
ASN
B
130
−76.934
−55.800
−33.646
1.00
49.37
C


ATOM
10017
O
ASN
B
130
−78.083
−55.456
−33.366
1.00
49.29
O


ATOM
10019
N
LEU
B
131
−75.868
−55.336
−32.996
1.00
48.98
N


ATOM
10020
CA
LEU
B
131
−75.957
−54.279
−31.988
1.00
48.56
C


ATOM
10022
CB
LEU
B
131
−74.797
−54.392
−30.991
1.00
48.44
C


ATOM
10025
CG
LEU
B
131
−74.759
−55.671
−30.148
1.00
48.19
C


ATOM
10027
CD1
LEU
B
131
−73.382
−55.852
−29.523
1.00
47.23
C


ATOM
10031
CD2
LEU
B
131
−75.861
−55.670
−29.083
1.00
47.77
C


ATOM
10035
C
LEU
B
131
−75.968
−52.875
−32.604
1.00
48.29
C


ATOM
10036
O
LEU
B
131
−76.022
−51.895
−31.874
1.00
48.44
O


ATOM
10038
N
LYS
B
132
−75.927
−52.771
−33.934
1.00
47.95
N


ATOM
10039
CA
LYS
B
132
−76.020
−51.471
−34.621
1.00
47.64
C


ATOM
10041
CB
LYS
B
132
−75.548
−51.590
−36.080
1.00
47.63
C


ATOM
10044
CG
LYS
B
132
−76.595
−52.235
−36.998
1.00
48.19
C


ATOM
10047
CD
LYS
B
132
−76.111
−52.483
−38.429
1.00
48.16
C


ATOM
10050
CE
LYS
B
132
−77.239
−53.040
−39.290
1.00
47.23
C


ATOM
10053
NZ
LYS
B
132
−76.725
−53.936
−40.331
1.00
47.01
N


ATOM
10057
C
LYS
B
132
−77.449
−50.907
−34.597
1.00
47.13
C


ATOM
10058
O
LYS
B
132
−77.683
−49.772
−35.005
1.00
46.87
O


ATOM
10060
N
GLU
B
133
−78.403
−51.716
−34.149
1.00
46.77
N


ATOM
10061
CA
GLU
B
133
−79.806
−51.315
−34.107
1.00
46.49
C


ATOM
10063
CB
GLU
B
133
−80.691
−52.458
−34.622
1.00
46.56
C


ATOM
10066
CG
GLU
B
133
−80.732
−52.515
−36.155
1.00
47.01
C


ATOM
10069
CD
GLU
B
133
−80.737
−53.926
−36.715
1.00
47.40
C


ATOM
10070
OE1
GLU
B
133
−81.500
−54.775
−36.206
1.00
47.97
O


ATOM
10071
OE2
GLU
B
133
−79.983
−54.177
−37.679
1.00
47.22
O


ATOM
10072
C
GLU
B
133
−80.260
−50.830
−32.723
1.00
45.92
C


ATOM
10073
O
GLU
B
133
−81.369
−50.308
−32.600
1.00
46.14
O


ATOM
10075
N
ASP
B
134
−79.424
−51.002
−31.691
1.00
44.98
N


ATOM
10076
CA
ASP
B
134
−79.611
−50.277
−30.428
1.00
44.14
C


ATOM
10078
CB
ASP
B
134
−79.375
−51.162
−29.197
1.00
43.95
C


ATOM
10081
CG
ASP
B
134
−79.646
−50.419
−27.883
1.00
43.57
C


ATOM
10082
OD1
ASP
B
134
−80.014
−49.230
−27.919
1.00
42.21
O


ATOM
10083
OD2
ASP
B
134
−79.488
−51.011
−26.802
1.00
43.69
O


ATOM
10084
C
ASP
B
134
−78.662
−49.073
−30.426
1.00
43.47
C


ATOM
10085
O
ASP
B
134
−77.478
−49.196
−30.104
1.00
43.55
O


ATOM
10087
N
ILE
B
135
−79.198
−47.908
−30.776
1.00
42.47
N


ATOM
10088
CA
ILE
B
135
−78.375
−46.739
−31.033
1.00
41.65
C


ATOM
10090
CB
ILE
B
135
−79.080
−45.783
−32.012
1.00
41.45
C


ATOM
10092
CG1
ILE
B
135
−78.877
−46.313
−33.434
1.00
41.74
C


ATOM
10095
CD1
ILE
B
135
−79.174
−45.331
−34.540
1.00
42.77
C


ATOM
10099
CG2
ILE
B
135
−78.533
−44.391
−31.905
1.00
41.68
C


ATOM
10103
C
ILE
B
135
−77.901
−46.059
−29.746
1.00
41.14
C


ATOM
10104
O
ILE
B
135
−76.752
−45.624
−29.676
1.00
41.11
O


ATOM
10106
N
LYS
B
136
−78.742
−45.998
−28.714
1.00
40.53
N


ATOM
10107
CA
LYS
B
136
−78.281
−45.480
−27.409
1.00
40.10
C


ATOM
10109
CB
LYS
B
136
−79.431
−45.361
−26.375
1.00
40.40
C


ATOM
10112
CG
LYS
B
136
−79.910
−46.697
−25.786
1.00
42.49
C


ATOM
10115
CD
LYS
B
136
−81.002
−46.574
−24.698
1.00
44.79
C


ATOM
10118
CE
LYS
B
136
−81.599
−47.994
−24.363
1.00
46.03
C


ATOM
10121
NZ
LYS
B
136
−82.231
−48.149
−22.994
1.00
46.23
N


ATOM
10125
C
LYS
B
136
−77.104
−46.327
−26.863
1.00
38.85
C


ATOM
10126
O
LYS
B
136
−76.246
−45.806
−26.143
1.00
38.91
O


ATOM
10128
N
ALA
B
137
−77.062
−47.617
−27.219
1.00
37.27
N


ATOM
10129
CA
ALA
B
137
−75.935
−48.486
−26.864
1.00
35.88
C


ATOM
10131
CB
ALA
B
137
−76.255
−49.952
−27.116
1.00
35.69
C


ATOM
10135
C
ALA
B
137
−74.708
−48.085
−27.645
1.00
34.63
C


ATOM
10136
O
ALA
B
137
−73.647
−47.894
−27.055
1.00
34.70
O


ATOM
10138
N
ILE
B
138
−74.845
−47.946
−28.965
1.00
33.12
N


ATOM
10139
CA
ILE
B
138
−73.693
−47.597
−29.805
1.00
31.97
C


ATOM
10141
CB
ILE
B
138
−74.020
−47.572
−31.296
1.00
31.50
C


ATOM
10143
CG1
ILE
B
138
−74.433
−48.955
−31.774
1.00
31.28
C


ATOM
10146
CD1
ILE
B
138
−73.460
−50.052
−31.417
1.00
30.86
C


ATOM
10150
CG2
ILE
B
138
−72.819
−47.138
−32.073
1.00
30.61
C


ATOM
10154
C
ILE
B
138
−73.124
−46.245
−29.413
1.00
31.57
C


ATOM
10155
O
ILE
B
138
−71.909
−46.090
−29.305
1.00
31.77
O


ATOM
10157
N
LEU
B
139
−73.997
−45.268
−29.192
1.00
30.92
N


ATOM
10158
CA
LEU
B
139
−73.555
−43.962
−28.713
1.00
30.29
C


ATOM
10160
CB
LEU
B
139
−74.732
−43.000
−28.530
1.00
30.13
C


ATOM
10163
CG
LEU
B
139
−74.778
−41.890
−29.574
1.00
29.96
C


ATOM
10165
CD1
LEU
B
139
−73.496
−41.060
−29.508
1.00
28.26
C


ATOM
10169
CD2
LEU
B
139
−76.022
−41.019
−29.397
1.00
29.48
C


ATOM
10173
C
LEU
B
139
−72.835
−44.136
−27.397
1.00
29.95
C


ATOM
10174
O
LEU
B
139
−71.709
−43.687
−27.232
1.00
30.45
O


ATOM
10176
N
SER
B
140
−73.492
−44.807
−26.463
1.00
29.33
N


ATOM
10177
CA
SER
B
140
−72.916
−45.061
−25.150
1.00
28.84
C


ATOM
10179
CB
SER
B
140
−73.900
−45.878
−24.298
1.00
28.93
C


ATOM
10182
OG
SER
B
140
−73.527
−45.870
−22.930
1.00
30.33
O


ATOM
10184
C
SER
B
140
−71.555
−45.770
−25.243
1.00
27.78
C


ATOM
10185
O
SER
B
140
−70.637
−45.454
−24.489
1.00
27.46
O


ATOM
10187
N
LEU
B
141
−71.432
−46.719
−26.167
1.00
26.82
N


ATOM
10188
CA
LEU
B
141
−70.178
−47.433
−26.348
1.00
26.34
C


ATOM
10190
CB
LEU
B
141
−70.366
−48.658
−27.256
1.00
26.16
C


ATOM
10193
CG
LEU
B
141
−69.098
−49.472
−27.584
1.00
26.09
C


ATOM
10195
CD1
LEU
B
141
−68.355
−49.960
−26.330
1.00
24.64
C


ATOM
10199
CD2
LEU
B
141
−69.455
−50.643
−28.481
1.00
25.94
C


ATOM
10203
C
LEU
B
141
−69.124
−46.476
−26.914
1.00
25.89
C


ATOM
10204
O
LEU
B
141
−68.025
−46.346
−26.366
1.00
25.77
O


ATOM
10206
N
TYR
B
142
−69.471
−45.809
−28.008
1.00
25.41
N


ATOM
10207
CA
TYR
B
142
−68.609
−44.804
−28.610
1.00
25.03
C


ATOM
10209
CB
TYR
B
142
−69.399
−43.982
−29.617
1.00
24.80
C


ATOM
10212
CG
TYR
B
142
−68.736
−42.695
−30.043
1.00
24.47
C


ATOM
10213
CD1
TYR
B
142
−67.761
−42.686
−31.029
1.00
24.18
C


ATOM
10215
CE1
TYR
B
142
−67.166
−41.516
−31.438
1.00
24.70
C


ATOM
10217
CZ
TYR
B
142
−67.548
−40.315
−30.871
1.00
25.80
C


ATOM
10218
OH
TYR
B
142
−66.940
−39.129
−31.279
1.00
27.63
O


ATOM
10220
CE2
TYR
B
142
−68.522
−40.299
−29.890
1.00
25.57
C


ATOM
10222
CD2
TYR
B
142
−69.110
−41.485
−29.485
1.00
24.73
C


ATOM
10224
C
TYR
B
142
−68.095
−43.883
−27.537
1.00
25.17
C


ATOM
10225
O
TYR
B
142
−66.886
−43.732
−27.360
1.00
25.40
O


ATOM
10227
N
GLU
B
143
−69.038
−43.293
−26.807
1.00
25.12
N


ATOM
10228
CA
GLU
B
143
−68.744
−42.256
−25.828
1.00
25.10
C


ATOM
10230
CB
GLU
B
143
−70.046
−41.726
−25.226
1.00
25.33
C


ATOM
10233
CG
GLU
B
143
−70.006
−40.244
−24.849
1.00
27.57
C


ATOM
10236
CD
GLU
B
143
−70.321
−39.313
−26.023
1.00
30.43
C


ATOM
10237
OE1
GLU
B
143
−69.343
−38.764
−26.583
1.00
32.89
O


ATOM
10238
OE2
GLU
B
143
−71.526
−39.130
−26.377
1.00
30.86
O


ATOM
10239
C
GLU
B
143
−67.794
−42.746
−24.729
1.00
24.43
C


ATOM
10240
O
GLU
B
143
−67.026
−41.959
−24.179
1.00
24.06
O


ATOM
10242
N
ALA
B
144
−67.840
−44.050
−24.449
1.00
24.01
N


ATOM
10243
CA
ALA
B
144
−67.033
−44.681
−23.399
1.00
23.74
C


ATOM
10245
CB
ALA
B
144
−67.726
−45.943
−22.883
1.00
23.48
C


ATOM
10249
C
ALA
B
144
−65.631
−45.025
−23.864
1.00
23.57
C


ATOM
10250
O
ALA
B
144
−64.720
−45.171
−23.060
1.00
23.69
O


ATOM
10252
N
SER
B
145
−65.455
−45.168
−25.165
1.00
23.45
N


ATOM
10253
CA
SER
B
145
−64.168
−45.568
−25.701
1.00
23.44
C


ATOM
10255
CB
SER
B
145
−64.300
−45.856
−27.214
1.00
23.63
C


ATOM
10258
OG
SER
B
145
−64.822
−44.748
−27.952
1.00
24.24
O


ATOM
10260
C
SER
B
145
−63.065
−44.520
−25.416
1.00
23.24
C


ATOM
10261
O
SER
B
145
−61.877
−44.848
−25.288
1.00
23.25
O


ATOM
10263
N
PHE
B
146
−63.454
−43.257
−25.285
1.00
22.92
N


ATOM
10264
CA
PHE
B
146
−62.464
−42.189
−25.170
1.00
22.35
C


ATOM
10266
CB
PHE
B
146
−63.090
−40.837
−25.478
1.00
22.09
C


ATOM
10269
CG
PHE
B
146
−63.505
−40.717
−26.911
1.00
22.40
C


ATOM
10270
CD1
PHE
B
146
−62.619
−40.229
−27.868
1.00
21.98
C


ATOM
10272
CE1
PHE
B
146
−62.985
−40.156
−29.201
1.00
21.80
C


ATOM
10274
CZ
PHE
B
146
−64.241
−40.579
−29.594
1.00
21.91
C


ATOM
10276
CE2
PHE
B
146
−65.123
−41.098
−28.649
1.00
22.29
C


ATOM
10278
CD2
PHE
B
146
−64.751
−41.170
−27.323
1.00
22.34
C


ATOM
10280
C
PHE
B
146
−61.762
−42.208
−23.837
1.00
21.91
C


ATOM
10281
O
PHE
B
146
−60.669
−41.683
−23.733
1.00
22.30
O


ATOM
10283
N
LEU
B
147
−62.361
−42.864
−22.841
1.00
21.27
N


ATOM
10284
CA
LEU
B
147
−61.727
−43.050
−21.535
1.00
20.49
C


ATOM
10286
CB
LEU
B
147
−62.791
−43.333
−20.478
1.00
20.14
C


ATOM
10289
CG
LEU
B
147
−63.609
−42.109
−20.102
1.00
18.87
C


ATOM
10291
CD1
LEU
B
147
−64.813
−42.020
−20.959
1.00
15.94
C


ATOM
10295
CD2
LEU
B
147
−63.988
−42.189
−18.642
1.00
18.62
C


ATOM
10299
C
LEU
B
147
−60.678
−44.163
−21.498
1.00
20.41
C


ATOM
10300
O
LEU
B
147
−60.243
−44.552
−20.415
1.00
20.44
O


ATOM
10302
N
ALA
B
148
−60.268
−44.664
−22.662
1.00
20.28
N


ATOM
10303
CA
ALA
B
148
−59.330
−45.784
−22.745
1.00
20.33
C


ATOM
10305
CB
ALA
B
148
−59.188
−46.231
−24.194
1.00
20.15
C


ATOM
10309
C
ALA
B
148
−57.952
−45.462
−22.170
1.00
20.54
C


ATOM
10310
O
ALA
B
148
−57.435
−44.366
−22.341
1.00
20.23
O


ATOM
10312
N
LEU
B
149
−57.366
−46.435
−21.486
1.00
21.19
N


ATOM
10313
CA
LEU
B
149
−55.969
−46.354
−21.067
1.00
22.02
C


ATOM
10315
CB
LEU
B
149
−55.768
−47.073
−19.725
1.00
21.90
C


ATOM
10318
CG
LEU
B
149
−56.541
−46.496
−18.523
1.00
21.50
C


ATOM
10320
CD1
LEU
B
149
−55.979
−47.006
−17.223
1.00
20.96
C


ATOM
10324
CD2
LEU
B
149
−56.520
−44.974
−18.505
1.00
20.95
C


ATOM
10328
C
LEU
B
149
−55.054
−46.930
−22.159
1.00
22.84
C


ATOM
10329
O
LEU
B
149
−55.507
−47.678
−23.017
1.00
22.91
O


ATOM
10331
N
GLU
B
150
−53.781
−46.550
−22.163
1.00
23.93
N


ATOM
10332
CA
GLU
B
150
−52.858
−47.071
−23.174
1.00
25.03
C


ATOM
10334
CB
GLU
B
150
−51.472
−46.431
−23.012
1.00
25.41
C


ATOM
10337
CG
GLU
B
150
−50.530
−46.608
−24.213
1.00
27.44
C


ATOM
10340
CD
GLU
B
150
−49.132
−45.987
−23.992
1.00
30.04
C


ATOM
10341
OE1
GLU
B
150
−48.925
−45.330
−22.943
1.00
31.41
O


ATOM
10342
OE2
GLU
B
150
−48.242
−46.161
−24.867
1.00
30.34
O


ATOM
10343
C
GLU
B
150
−52.787
−48.600
−23.024
1.00
25.36
C


ATOM
10344
O
GLU
B
150
−52.613
−49.098
−21.911
1.00
25.58
O


ATOM
10346
N
GLY
B
151
−52.968
−49.337
−24.122
1.00
25.79
N


ATOM
10347
CA
GLY
B
151
−52.959
−50.816
−24.086
1.00
26.11
C


ATOM
10350
C
GLY
B
151
−54.323
−51.522
−24.047
1.00
26.44
C


ATOM
10351
O
GLY
B
151
−54.393
−52.747
−24.135
1.00
26.91
O


ATOM
10353
N
GLU
B
152
−55.407
−50.768
−23.899
1.00
26.58
N


ATOM
10354
CA
GLU
B
152
−56.757
−51.319
−23.971
1.00
26.57
C


ATOM
10356
CB
GLU
B
152
−57.697
−50.531
−23.065
1.00
26.74
C


ATOM
10359
CG
GLU
B
152
−57.291
−50.595
−21.596
1.00
27.63
C


ATOM
10362
CD
GLU
B
152
−58.271
−49.893
−20.668
1.00
28.86
C


ATOM
10363
OE1
GLU
B
152
−58.239
−50.192
−19.449
1.00
28.80
O


ATOM
10364
OE2
GLU
B
152
−59.066
−49.044
−21.154
1.00
29.26
O


ATOM
10365
C
GLU
B
152
−57.254
−51.303
−25.415
1.00
26.53
C


ATOM
10366
O
GLU
B
152
−57.937
−50.381
−25.869
1.00
26.00
O


ATOM
10368
N
ASN
B
153
−56.888
−52.354
−26.127
1.00
26.77
N


ATOM
10369
CA
ASN
B
153
−57.189
−52.498
−27.545
1.00
27.05
C


ATOM
10371
CB
ASN
B
153
−56.345
−53.640
−28.104
1.00
27.12
C


ATOM
10374
CG
ASN
B
153
−56.770
−54.992
−27.549
1.00
27.81
C


ATOM
10375
OD1
ASN
B
153
−56.540
−55.307
−26.379
1.00
27.65
O


ATOM
10376
ND2
ASN
B
153
−57.426
−55.782
−28.382
1.00
29.43
N


ATOM
10379
C
ASN
B
153
−58.659
−52.788
−27.873
1.00
27.09
C


ATOM
10380
O
ASN
B
153
−59.053
−52.658
−29.029
1.00
27.11
O


ATOM
10382
N
ILE
B
154
−59.448
−53.223
−26.884
1.00
27.17
N


ATOM
10383
CA
ILE
B
154
−60.865
−53.544
−27.100
1.00
27.11
C


ATOM
10385
CB
ILE
B
154
−61.454
−54.369
−25.956
1.00
26.99
C


ATOM
10387
CG1
ILE
B
154
−60.811
−55.750
−25.907
1.00
27.16
C


ATOM
10390
CD1
ILE
B
154
−61.309
−56.622
−24.746
1.00
27.41
C


ATOM
10394
CG2
ILE
B
154
−62.953
−54.519
−26.124
1.00
26.65
C


ATOM
10398
C
ILE
B
154
−61.704
−52.284
−27.229
1.00
27.43
C


ATOM
10399
O
ILE
B
154
−62.718
−52.279
−27.939
1.00
27.39
O


ATOM
10401
N
LEU
B
155
−61.296
−51.226
−26.522
1.00
27.76
N


ATOM
10402
CA
LEU
B
155
−61.964
−49.918
−26.609
1.00
27.75
C


ATOM
10404
CB
LEU
B
155
−61.578
−49.024
−25.430
1.00
27.31
C


ATOM
10407
CG
LEU
B
155
−61.968
−49.544
−24.052
1.00
26.64
C


ATOM
10409
CD1
LEU
B
155
−61.444
−48.631
−22.981
1.00
25.82
C


ATOM
10413
CD2
LEU
B
155
−63.465
−49.674
−23.944
1.00
26.45
C


ATOM
10417
C
LEU
B
155
−61.620
−49.226
−27.924
1.00
28.32
C


ATOM
10418
O
LEU
B
155
−62.499
−48.672
−28.583
1.00
28.72
O


ATOM
10420
N
ASP
B
156
−60.348
−49.264
−28.309
1.00
28.84
N


ATOM
10421
CA
ASP
B
156
−59.934
−48.737
−29.601
1.00
29.43
C


ATOM
10423
CB
ASP
B
156
−58.423
−48.928
−29.823
1.00
29.92
C


ATOM
10426
CG
ASP
B
156
−57.566
−47.912
−29.045
1.00
31.35
C


ATOM
10427
OD1
ASP
B
156
−57.803
−46.680
−29.162
1.00
33.23
O


ATOM
10428
OD2
ASP
B
156
−56.640
−48.351
−28.324
1.00
33.08
O


ATOM
10429
C
ASP
B
156
−60.719
−49.411
−30.717
1.00
29.36
C


ATOM
10430
O
ASP
B
156
−61.075
−48.768
−31.697
1.00
29.33
O


ATOM
10432
N
GLU
B
157
−60.981
−50.705
−30.558
1.00
29.63
N


ATOM
10433
CA
GLU
B
157
−61.845
−51.457
−31.479
1.00
29.98
C


ATOM
10435
CB
GLU
B
157
−61.729
−52.971
−31.234
1.00
30.30
C


ATOM
10438
CG
GLU
B
157
−60.664
−53.647
−32.087
1.00
31.79
C


ATOM
10441
CD
GLU
B
157
−60.075
−54.901
−31.439
1.00
33.87
C


ATOM
10442
OE1
GLU
B
157
−60.793
−55.560
−30.640
1.00
35.07
O


ATOM
10443
OE2
GLU
B
157
−58.894
−55.223
−31.743
1.00
33.70
O


ATOM
10444
C
GLU
B
157
−63.304
−51.030
−31.349
1.00
29.53
C


ATOM
10445
O
GLU
B
157
−63.999
−50.878
−32.351
1.00
29.53
O


ATOM
10447
N
ALA
B
158
−63.758
−50.853
−30.112
1.00
29.11
N


ATOM
10448
CA
ALA
B
158
−65.104
−50.370
−29.840
1.00
28.87
C


ATOM
10450
CB
ALA
B
158
−65.307
−50.195
−28.351
1.00
28.82
C


ATOM
10454
C
ALA
B
158
−65.383
−49.065
−30.567
1.00
28.73
C


ATOM
10455
O
ALA
B
158
−66.485
−48.857
−31.065
1.00
28.80
O


ATOM
10457
N
LYS
B
159
−64.385
−48.197
−30.642
1.00
28.65
N


ATOM
10458
CA
LYS
B
159
−64.553
−46.910
−31.296
1.00
28.98
C


ATOM
10460
CB
LYS
B
159
−63.439
−45.950
−30.857
1.00
29.16
C


ATOM
10463
CG
LYS
B
159
−63.558
−44.536
−31.426
1.00
29.82
C


ATOM
10466
CD
LYS
B
159
−62.812
−43.493
−30.592
1.00
30.96
C


ATOM
10469
CE
LYS
B
159
−61.295
−43.610
−30.691
1.00
31.57
C


ATOM
10472
NZ
LYS
B
159
−60.630
−42.426
−30.079
1.00
31.49
N


ATOM
10476
C
LYS
B
159
−64.594
−47.041
−32.826
1.00
29.08
C


ATOM
10477
O
LYS
B
159
−65.385
−46.372
−33.486
1.00
28.73
O


ATOM
10479
N
VAL
B
160
−63.736
−47.891
−33.386
1.00
29.53
N


ATOM
10480
CA
VAL
B
160
−63.686
−48.087
−34.836
1.00
29.89
C


ATOM
10482
CB
VAL
B
160
−62.466
−48.971
−35.282
1.00
29.93
C


ATOM
10484
CG1
VAL
B
160
−62.576
−49.375
−36.756
1.00
29.58
C


ATOM
10488
CG2
VAL
B
160
−61.152
−48.244
−35.040
1.00
29.41
C


ATOM
10492
C
VAL
B
160
−65.001
−48.723
−35.263
1.00
30.32
C


ATOM
10493
O
VAL
B
160
−65.507
−48.459
−36.362
1.00
30.41
O


ATOM
10495
N
PHE
B
161
−65.558
−49.540
−34.371
1.00
30.83
N


ATOM
10496
CA
PHE
B
161
−66.854
−50.178
−34.599
1.00
31.34
C


ATOM
10498
CB
PHE
B
161
−67.090
−51.321
−33.599
1.00
31.34
C


ATOM
10501
CG
PHE
B
161
−68.492
−51.834
−33.603
1.00
31.09
C


ATOM
10502
CD1
PHE
B
161
−68.940
−52.636
−34.633
1.00
31.61
C


ATOM
10504
CE1
PHE
B
161
−70.249
−53.098
−34.649
1.00
31.48
C


ATOM
10506
CZ
PHE
B
161
−71.114
−52.748
−33.628
1.00
30.76
C


ATOM
10508
CE2
PHE
B
161
−70.677
−51.944
−32.606
1.00
30.19
C


ATOM
10510
CD2
PHE
B
161
−69.378
−51.486
−32.597
1.00
30.70
C


ATOM
10512
C
PHE
B
161
−67.992
−49.173
−34.504
1.00
31.65
C


ATOM
10513
O
PHE
B
161
−68.785
−49.038
−35.432
1.00
31.62
O


ATOM
10515
N
ALA
B
162
−68.068
−48.483
−33.373
1.00
32.26
N


ATOM
10516
CA
ALA
B
162
−69.135
−47.519
−33.129
1.00
32.84
C


ATOM
10518
CB
ALA
B
162
−68.948
−46.854
−31.778
1.00
32.66
C


ATOM
10522
C
ALA
B
162
−69.224
−46.474
−34.245
1.00
33.40
C


ATOM
10523
O
ALA
B
162
−70.229
−46.408
−34.937
1.00
33.45
O


ATOM
10525
N
ILE
B
163
−68.164
−45.697
−34.444
1.00
34.38
N


ATOM
10526
CA
ILE
B
163
−68.166
−44.610
−35.439
1.00
35.23
C


ATOM
10528
CB
ILE
B
163
−66.734
−44.062
−35.733
1.00
35.18
C


ATOM
10530
CG1
ILE
B
163
−66.092
−43.457
−34.488
1.00
35.21
C


ATOM
10533
CD1
ILE
B
163
−64.620
−43.171
−34.663
1.00
35.96
C


ATOM
10537
CG2
ILE
B
163
−66.778
−42.977
−36.799
1.00
34.72
C


ATOM
10541
C
ILE
B
163
−68.778
−45.052
−36.766
1.00
36.18
C


ATOM
10542
O
ILE
B
163
−69.588
−44.335
−37.360
1.00
35.87
O


ATOM
10544
N
SER
B
164
−68.379
−46.240
−37.217
1.00
37.67
N


ATOM
10545
CA
SER
B
164
−68.761
−46.752
−38.539
1.00
38.74
C


ATOM
10547
CB
SER
B
164
−68.104
−48.117
−38.815
1.00
38.80
C


ATOM
10550
OG
SER
B
164
−68.733
−49.158
−38.077
1.00
39.07
O


ATOM
10552
C
SER
B
164
−70.277
−46.861
−38.699
1.00
39.51
C


ATOM
10553
O
SER
B
164
−70.820
−46.494
−39.737
1.00
39.96
O


ATOM
10555
N
HIS
B
165
−70.962
−47.363
−37.680
1.00
40.28
N


ATOM
10556
CA
HIS
B
165
−72.410
−47.485
−37.767
1.00
41.07
C


ATOM
10558
CB
HIS
B
165
−72.911
−48.686
−36.957
1.00
41.38
C


ATOM
10561
CG
HIS
B
165
−72.571
−50.005
−37.587
1.00
42.65
C


ATOM
10562
ND1
HIS
B
165
−71.647
−50.875
−37.046
1.00
43.54
N


ATOM
10564
CE1
HIS
B
165
−71.535
−51.938
−37.825
1.00
43.54
C


ATOM
10566
NE2
HIS
B
165
−72.346
−51.785
−38.858
1.00
43.25
N


ATOM
10568
CD2
HIS
B
165
−73.001
−50.582
−38.737
1.00
43.31
C


ATOM
10570
C
HIS
B
165
−73.125
−46.193
−37.380
1.00
41.21
C


ATOM
10571
O
HIS
B
165
−74.257
−45.980
−37.805
1.00
41.53
O


ATOM
10573
N
LEU
B
166
−72.459
−45.326
−36.614
1.00
41.32
N


ATOM
10574
CA
LEU
B
166
−73.042
−44.039
−36.201
1.00
41.31
C


ATOM
10576
CB
LEU
B
166
−72.317
−43.460
−34.973
1.00
41.14
C


ATOM
10579
CG
LEU
B
166
−72.732
−43.975
−33.589
1.00
40.03
C


ATOM
10581
CD1
LEU
B
166
−71.673
−43.634
−32.579
1.00
38.86
C


ATOM
10585
CD2
LEU
B
166
−74.077
−43.421
−33.151
1.00
38.52
C


ATOM
10589
C
LEU
B
166
−73.045
−42.992
−37.313
1.00
41.80
C


ATOM
10590
O
LEU
B
166
−74.006
−42.230
−37.442
1.00
41.73
O


ATOM
10592
N
LYS
B
167
−71.982
−42.947
−38.116
1.00
42.47
N


ATOM
10593
CA
LYS
B
167
−71.847
−41.891
−39.137
1.00
43.00
C


ATOM
10595
CB
LYS
B
167
−70.416
−41.822
−39.702
1.00
43.12
C


ATOM
10598
CG
LYS
B
167
−69.937
−43.068
−40.450
1.00
44.12
C


ATOM
10601
CD
LYS
B
167
−69.138
−42.725
−41.736
1.00
45.36
C


ATOM
10604
CE
LYS
B
167
−67.898
−41.842
−41.483
1.00
45.65
C


ATOM
10607
NZ
LYS
B
167
−67.267
−41.400
−42.762
1.00
44.94
N


ATOM
10611
C
LYS
B
167
−72.873
−41.950
−40.283
1.00
42.99
C


ATOM
10612
O
LYS
B
167
−72.826
−41.124
−41.182
1.00
42.95
O


ATOM
10614
N
GLU
B
168
−73.791
−42.912
−40.244
1.00
43.23
N


ATOM
10615
CA
GLU
B
168
−74.928
−42.940
−41.167
1.00
43.44
C


ATOM
10617
CB
GLU
B
168
−74.652
−43.969
−42.272
1.00
43.58
C


ATOM
10620
CG
GLU
B
168
−73.918
−43.363
−43.501
1.00
44.36
C


ATOM
10623
CD
GLU
B
168
−72.618
−44.082
−43.895
1.00
44.53
C


ATOM
10624
OE1
GLU
B
168
−71.853
−44.494
−42.992
1.00
44.71
O


ATOM
10625
OE2
GLU
B
168
−72.351
−44.197
−45.115
1.00
43.31
O


ATOM
10626
C
GLU
B
168
−76.263
−43.214
−40.442
1.00
43.24
C


ATOM
10627
O
GLU
B
168
−76.932
−42.291
−39.942
1.00
42.54
O


ATOM
10629
N
GLY
B
175
−83.548
−42.037
−36.239
1.00
49.43
N


ATOM
10630
CA
GLY
B
175
−84.473
−41.266
−35.416
1.00
49.54
C


ATOM
10633
C
GLY
B
175
−84.709
−39.884
−36.001
1.00
49.76
C


ATOM
10634
O
GLY
B
175
−84.828
−39.741
−37.220
1.00
49.70
O


ATOM
10636
N
LYS
B
176
−84.766
−38.870
−35.131
1.00
49.95
N


ATOM
10637
CA
LYS
B
176
−85.014
−37.468
−35.534
1.00
50.05
C


ATOM
10639
CB
LYS
B
176
−86.522
−37.225
−35.730
1.00
50.27
C


ATOM
10642
CG
LYS
B
176
−87.301
−36.813
−34.460
1.00
51.33
C


ATOM
10645
CD
LYS
B
176
−88.553
−37.658
−34.202
1.00
52.21
C


ATOM
10648
CE
LYS
B
176
−88.841
−37.754
−32.695
1.00
52.36
C


ATOM
10651
NZ
LYS
B
176
−90.029
−38.607
−32.422
1.00
52.15
N


ATOM
10655
C
LYS
B
176
−84.427
−36.432
−34.554
1.00
49.75
C


ATOM
10656
O
LYS
B
176
−83.990
−35.370
−34.972
1.00
49.88
O


ATOM
10658
N
GLU
B
177
−84.473
−36.729
−33.254
1.00
49.46
N


ATOM
10659
CA
GLU
B
177
−83.769
−35.959
−32.221
1.00
48.92
C


ATOM
10661
CB
GLU
B
177
−84.628
−35.813
−30.946
1.00
49.03
C


ATOM
10664
CG
GLU
B
177
−84.196
−36.691
−29.732
1.00
49.60
C


ATOM
10667
CD
GLU
B
177
−85.278
−36.871
−28.662
1.00
50.10
C


ATOM
10668
OE1
GLU
B
177
−84.998
−37.576
−27.670
1.00
49.78
O


ATOM
10669
OE2
GLU
B
177
−86.401
−36.336
−28.808
1.00
50.78
O


ATOM
10670
C
GLU
B
177
−82.469
−36.703
−31.924
1.00
48.19
C


ATOM
10671
O
GLU
B
177
−81.421
−36.085
−31.721
1.00
48.68
O


ATOM
10673
N
LEU
B
178
−82.557
−38.037
−31.900
1.00
46.98
N


ATOM
10674
CA
LEU
B
178
−81.398
−38.924
−31.817
1.00
45.91
C


ATOM
10676
CB
LEU
B
178
−81.838
−40.387
−31.945
1.00
45.70
C


ATOM
10679
CG
LEU
B
178
−81.111
−41.408
−31.074
1.00
45.24
C


ATOM
10681
CD1
LEU
B
178
−81.498
−41.225
−29.609
1.00
44.88
C


ATOM
10685
CD2
LEU
B
178
−81.413
−42.826
−31.539
1.00
44.19
C


ATOM
10689
C
LEU
B
178
−80.423
−38.571
−32.934
1.00
45.27
C


ATOM
10690
O
LEU
B
178
−79.211
−38.624
−32.759
1.00
44.99
O


ATOM
10692
N
ALA
B
179
−80.970
−38.203
−34.088
1.00
44.75
N


ATOM
10693
CA
ALA
B
179
−80.174
−37.661
−35.181
1.00
44.24
C


ATOM
10695
CB
ALA
B
179
−81.088
−37.069
−36.231
1.00
44.30
C


ATOM
10699
C
ALA
B
179
−79.192
−36.602
−34.683
1.00
43.65
C


ATOM
10700
O
ALA
B
179
−78.028
−36.604
−35.061
1.00
43.56
O


ATOM
10702
N
GLU
B
180
−79.675
−35.707
−33.827
1.00
43.01
N


ATOM
10703
CA
GLU
B
180
−78.863
−34.610
−33.311
1.00
42.49
C


ATOM
10705
CB
GLU
B
180
−79.749
−33.477
−32.789
1.00
42.85
C


ATOM
10708
CG
GLU
B
180
−80.574
−32.800
−33.881
1.00
44.23
C


ATOM
10711
CD
GLU
B
180
−80.991
−31.382
−33.519
1.00
45.80
C


ATOM
10712
OE1
GLU
B
180
−80.098
−30.580
−33.156
1.00
46.69
O


ATOM
10713
OE2
GLU
B
180
−82.203
−31.065
−33.610
1.00
46.71
O


ATOM
10714
C
GLU
B
180
−77.944
−35.075
−32.210
1.00
41.39
C


ATOM
10715
O
GLU
B
180
−76.843
−34.562
−32.071
1.00
41.27
O


ATOM
10717
N
GLN
B
181
−78.404
−36.035
−31.419
1.00
40.20
N


ATOM
10718
CA
GLN
B
181
−77.570
−36.632
−30.394
1.00
39.45
C


ATOM
10720
CB
GLN
B
181
−78.280
−37.809
−29.736
1.00
39.88
C


ATOM
10723
CG
GLN
B
181
−78.005
−37.959
−28.249
1.00
41.64
C


ATOM
10726
CD
GLN
B
181
−78.873
−37.036
−27.404
1.00
44.00
C


ATOM
10727
OE1
GLN
B
181
−80.109
−36.992
−27.581
1.00
45.65
O


ATOM
10728
NE2
GLN
B
181
−78.235
−36.291
−26.474
1.00
43.41
N


ATOM
10731
C
GLN
B
181
−76.270
−37.120
−31.003
1.00
38.28
C


ATOM
10732
O
GLN
B
181
−75.203
−36.767
−30.523
1.00
38.27
O


ATOM
10734
N
VAL
B
182
−76.358
−37.911
−32.072
1.00
37.05
N


ATOM
10735
CA
VAL
B
182
−75.160
−38.515
−32.675
1.00
36.20
C


ATOM
10737
CB
VAL
B
182
−75.486
−39.661
−33.681
1.00
36.11
C


ATOM
10739
CG1
VAL
B
182
−76.493
−40.641
−33.080
1.00
36.13
C


ATOM
10743
CG2
VAL
B
182
−75.977
−39.106
−35.004
1.00
36.06
C


ATOM
10747
C
VAL
B
182
−74.238
−37.495
−33.364
1.00
35.44
C


ATOM
10748
O
VAL
B
182
−73.012
−37.682
−33.375
1.00
35.19
O


ATOM
10750
N
SER
B
183
−74.811
−36.432
−33.936
1.00
34.41
N


ATOM
10751
CA
SER
B
183
−73.998
−35.416
−34.630
1.00
33.74
C


ATOM
10753
CB
SER
B
183
−74.858
−34.438
−35.422
1.00
33.58
C


ATOM
10756
OG
SER
B
183
−76.003
−35.085
−35.925
1.00
34.52
O


ATOM
10758
C
SER
B
183
−73.184
−34.644
−33.625
1.00
32.88
C


ATOM
10759
O
SER
B
183
−72.055
−34.251
−33.906
1.00
33.25
O


ATOM
10761
N
HIS
B
184
−73.788
−34.427
−32.461
1.00
31.83
N


ATOM
10762
CA
HIS
B
184
−73.150
−33.787
−31.326
1.00
30.98
C


ATOM
10764
CB
HIS
B
184
−74.188
−33.627
−30.212
1.00
31.14
C


ATOM
10767
CG
HIS
B
184
−73.710
−32.849
−29.030
1.00
31.91
C


ATOM
10768
ND1
HIS
B
184
−73.326
−31.527
−29.114
1.00
33.18
N


ATOM
10770
CE1
HIS
B
184
−72.960
−31.107
−27.914
1.00
33.10
C


ATOM
10772
NE2
HIS
B
184
−73.106
−32.102
−27.056
1.00
31.85
N


ATOM
10774
CD2
HIS
B
184
−73.583
−33.200
−27.727
1.00
31.86
C


ATOM
10776
C
HIS
B
184
−71.968
−34.632
−30.865
1.00
30.13
C


ATOM
10777
O
HIS
B
184
−70.863
−34.122
−30.709
1.00
29.95
O


ATOM
10779
N
ALA
B
185
−72.194
−35.931
−30.683
1.00
29.27
N


ATOM
10780
CA
ALA
B
185
−71.123
−36.856
−30.281
1.00
28.64
C


ATOM
10782
CB
ALA
B
185
−71.689
−38.233
−29.986
1.00
28.44
C


ATOM
10786
C
ALA
B
185
−70.017
−36.965
−31.330
1.00
28.08
C


ATOM
10787
O
ALA
B
185
−68.839
−37.043
−30.992
1.00
28.26
O


ATOM
10789
N
LEU
B
186
−70.394
−36.984
−32.602
1.00
27.40
N


ATOM
10790
CA
LEU
B
186
−69.412
−37.101
−33.674
1.00
26.84
C


ATOM
10792
CB
LEU
B
186
−70.088
−37.481
−35.000
1.00
26.75
C


ATOM
10795
CG
LEU
B
186
−70.085
−38.983
−35.320
1.00
26.70
C


ATOM
10797
CD1
LEU
B
186
−70.214
−39.868
−34.075
1.00
27.07
C


ATOM
10801
CD2
LEU
B
186
−71.179
−39.306
−36.303
1.00
26.29
C


ATOM
10805
C
LEU
B
186
−68.594
−35.822
−33.815
1.00
26.45
C


ATOM
10806
O
LEU
B
186
−67.449
−35.875
−34.237
1.00
26.48
O


ATOM
10808
N
GLU
B
187
−69.186
−34.685
−33.454
1.00
25.93
N


ATOM
10809
CA
GLU
B
187
−68.479
−33.406
−33.403
1.00
25.57
C


ATOM
10811
CB
GLU
B
187
−69.447
−32.310
−32.962
1.00
25.61
C


ATOM
10814
CG
GLU
B
187
−69.035
−30.899
−33.325
1.00
26.29
C


ATOM
10817
CD
GLU
B
187
−69.930
−29.857
−32.671
1.00
27.10
C


ATOM
10818
OE1
GLU
B
187
−70.312
−30.057
−31.487
1.00
26.51
O


ATOM
10819
OE2
GLU
B
187
−70.251
−28.846
−33.344
1.00
27.54
O


ATOM
10820
C
GLU
B
187
−67.307
−33.490
−32.418
1.00
25.13
C


ATOM
10821
O
GLU
B
187
−66.155
−33.165
−32.749
1.00
24.85
O


ATOM
10823
N
LEU
B
188
−67.625
−33.938
−31.204
1.00
24.58
N


ATOM
10824
CA
LEU
B
188
−66.644
−34.137
−30.148
1.00
24.06
C


ATOM
10826
CB
LEU
B
188
−66.343
−32.817
−29.451
1.00
24.12
C


ATOM
10829
CG
LEU
B
188
−65.042
−32.714
−28.670
1.00
23.61
C


ATOM
10831
CD1
LEU
B
188
−63.895
−32.872
−29.629
1.00
23.12
C


ATOM
10835
CD2
LEU
B
188
−64.976
−31.368
−27.964
1.00
23.18
C


ATOM
10839
C
LEU
B
188
−67.248
−35.087
−29.140
1.00
23.70
C


ATOM
10840
O
LEU
B
188
−68.392
−34.901
−28.743
1.00
23.68
O


ATOM
10842
N
PRO
B
189
−66.493
−36.106
−28.713
1.00
23.34
N


ATOM
10843
CA
PRO
B
189
−67.031
−36.983
−27.692
1.00
23.08
C


ATOM
10845
CB
PRO
B
189
−66.018
−38.115
−27.638
1.00
22.96
C


ATOM
10848
CG
PRO
B
189
−64.743
−37.452
−27.940
1.00
23.22
C


ATOM
10851
CD
PRO
B
189
−65.059
−36.350
−28.929
1.00
23.54
C


ATOM
10854
C
PRO
B
189
−67.053
−36.229
−26.387
1.00
22.86
C


ATOM
10855
O
PRO
B
189
−66.284
−35.285
−26.215
1.00
23.03
O


ATOM
10856
N
LEU
B
190
−67.912
−36.628
−25.465
1.00
22.56
N


ATOM
10857
CA
LEU
B
190
−68.152
−35.773
−24.323
1.00
22.41
C


ATOM
10859
CB
LEU
B
190
−69.567
−35.970
−23.766
1.00
23.04
C


ATOM
10862
CG
LEU
B
190
−69.853
−37.136
−22.848
1.00
23.41
C


ATOM
10864
CD1
LEU
B
190
−69.303
−36.733
−21.481
1.00
25.02
C


ATOM
10868
CD2
LEU
B
190
−71.342
−37.411
−22.820
1.00
21.69
C


ATOM
10872
C
LEU
B
190
−67.065
−35.899
−23.266
1.00
21.56
C


ATOM
10873
O
LEU
B
190
−66.860
−34.976
−22.477
1.00
21.67
O


ATOM
10875
N
HIS
B
191
−66.320
−36.998
−23.283
1.00
20.39
N


ATOM
10876
CA
HIS
B
191
−65.089
−37.037
−22.490
1.00
19.40
C


ATOM
10878
CB
HIS
B
191
−64.399
−38.393
−22.597
1.00
19.19
C


ATOM
10881
CG
HIS
B
191
−63.222
−38.530
−21.689
1.00
19.32
C


ATOM
10882
ND1
HIS
B
191
−63.347
−38.562
−20.317
1.00
20.95
N


ATOM
10884
CE1
HIS
B
191
−62.145
−38.659
−19.771
1.00
21.15
C


ATOM
10886
NE2
HIS
B
191
−61.245
−38.689
−20.741
1.00
19.36
N


ATOM
10888
CD2
HIS
B
191
−61.893
−38.604
−21.948
1.00
19.49
C


ATOM
10890
C
HIS
B
191
−64.094
−35.913
−22.841
1.00
18.62
C


ATOM
10891
O
HIS
B
191
−63.150
−35.677
−22.085
1.00
18.59
O


ATOM
10893
N
ARG
B
192
−64.307
−35.216
−23.957
1.00
17.72
N


ATOM
10894
CA
ARG
B
192
−63.394
−34.150
−24.394
1.00
17.57
C


ATOM
10896
CB
ARG
B
192
−62.838
−34.504
−25.773
1.00
17.81
C


ATOM
10899
CG
ARG
B
192
−61.971
−35.736
−25.781
1.00
18.59
C


ATOM
10902
CD
ARG
B
192
−61.484
−36.067
−27.181
1.00
20.10
C


ATOM
10905
NE
ARG
B
192
−60.462
−37.116
−27.165
1.00
21.62
N


ATOM
10907
CZ
ARG
B
192
−59.919
−37.648
−28.254
1.00
22.77
C


ATOM
10908
NH1
ARG
B
192
−60.304
−37.248
−29.454
1.00
24.20
N


ATOM
10911
NH2
ARG
B
192
−58.987
−38.581
−28.149
1.00
23.37
N


ATOM
10914
C
ARG
B
192
−63.983
−32.726
−24.440
1.00
17.01
C


ATOM
10915
O
ARG
B
192
−63.242
−31.739
−24.526
1.00
15.78
O


ATOM
10917
N
ARG
B
193
−65.309
−32.634
−24.392
1.00
17.01
N


ATOM
10918
CA
ARG
B
193
−66.015
−31.350
−24.453
1.00
16.99
C


ATOM
10920
CB
ARG
B
193
−67.476
−31.582
−24.892
1.00
17.29
C


ATOM
10923
CG
ARG
B
193
−68.192
−30.318
−25.377
1.00
18.42
C


ATOM
10926
CD
ARG
B
193
−69.664
−30.559
−25.646
1.00
19.63
C


ATOM
10929
NE
ARG
B
193
−69.912
−31.592
−26.648
1.00
21.16
N


ATOM
10931
CZ
ARG
B
193
−69.868
−31.398
−27.971
1.00
23.56
C


ATOM
10932
NH1
ARG
B
193
−69.575
−30.210
−28.496
1.00
23.37
N


ATOM
10935
NH2
ARG
B
193
−70.112
−32.413
−28.789
1.00
25.37
N


ATOM
10938
C
ARG
B
193
−65.974
−30.658
−23.087
1.00
16.46
C


ATOM
10939
O
ARG
B
193
−66.040
−31.323
−22.034
1.00
16.47
O


ATOM
10941
N
THR
B
194
−65.864
−29.335
−23.078
1.00
15.84
N


ATOM
10942
CA
THR
B
194
−65.886
−28.629
−21.799
1.00
15.67
C


ATOM
10944
CB
THR
B
194
−65.354
−27.199
−21.884
1.00
15.48
C


ATOM
10946
OG1
THR
B
194
−66.077
−26.478
−22.882
1.00
15.55
O


ATOM
10948
CG2
THR
B
194
−63.877
−27.189
−22.211
1.00
14.65
C


ATOM
10952
C
THR
B
194
−67.310
−28.613
−21.270
1.00
15.79
C


ATOM
10953
O
THR
B
194
−68.264
−28.652
−22.030
1.00
15.73
O


ATOM
10955
N
GLN
B
195
−67.444
−28.557
−19.957
1.00
16.20
N


ATOM
10956
CA
GLN
B
195
−68.737
−28.711
−19.328
1.00
16.84
C


ATOM
10958
CB
GLN
B
195
−68.601
−28.781
−17.816
1.00
17.07
C


ATOM
10961
CG
GLN
B
195
−69.921
−28.440
−17.153
1.00
18.86
C


ATOM
10964
CD
GLN
B
195
−69.999
−28.875
−15.746
1.00
20.95
C


ATOM
10965
OE1
GLN
B
195
−69.099
−29.511
−15.245
1.00
24.63
O


ATOM
10966
NE2
GLN
B
195
−71.071
−28.529
−15.082
1.00
22.36
N


ATOM
10969
C
GLN
B
195
−69.757
−27.623
−19.669
1.00
16.93
C


ATOM
10970
O
GLN
B
195
−70.858
−27.931
−20.136
1.00
16.98
O


ATOM
10972
N
ARG
B
196
−69.430
−26.366
−19.386
1.00
16.92
N


ATOM
10973
CA
ARG
B
196
−70.358
−25.301
−19.685
1.00
17.06
C


ATOM
10975
CB
ARG
B
196
−69.719
−23.932
−19.459
1.00
17.00
C


ATOM
10978
CG
ARG
B
196
−70.095
−23.289
−18.120
1.00
17.09
C


ATOM
10981
CD
ARG
B
196
−70.283
−24.320
−16.995
1.00
17.56
C


ATOM
10984
NE
ARG
B
196
−71.452
−24.043
−16.146
1.00
17.64
N


ATOM
10986
CZ
ARG
B
196
−72.289
−24.965
−15.664
1.00
17.93
C


ATOM
10987
NH1
ARG
B
196
−72.136
−26.244
−15.966
1.00
17.72
N


ATOM
10990
NH2
ARG
B
196
−73.305
−24.611
−14.878
1.00
18.27
N


ATOM
10993
C
ARG
B
196
−70.881
−25.481
−21.098
1.00
17.42
C


ATOM
10994
O
ARG
B
196
−72.079
−25.519
−21.317
1.00
17.66
O


ATOM
10996
N
LEU
B
197
−69.984
−25.676
−22.044
1.00
18.04
N


ATOM
10997
CA
LEU
B
197
−70.379
−25.925
−23.425
1.00
18.51
C


ATOM
10999
CB
LEU
B
197
−69.133
−26.085
−24.289
1.00
18.52
C


ATOM
11002
CG
LEU
B
197
−68.998
−25.185
−25.508
1.00
18.06
C


ATOM
11004
CD1
LEU
B
197
−68.209
−23.932
−25.205
1.00
15.52
C


ATOM
11008
CD2
LEU
B
197
−68.293
−26.002
−26.574
1.00
20.21
C


ATOM
11012
C
LEU
B
197
−71.290
−27.163
−23.580
1.00
19.14
C


ATOM
11013
O
LEU
B
197
−72.226
−27.150
−24.372
1.00
19.52
O


ATOM
11015
N
GLU
B
198
−71.030
−28.235
−22.842
1.00
19.72
N


ATOM
11016
CA
GLU
B
198
−71.918
−29.396
−22.909
1.00
20.43
C


ATOM
11018
CB
GLU
B
198
−71.272
−30.631
−22.270
1.00
20.65
C


ATOM
11021
CG
GLU
B
198
−72.176
−31.880
−22.172
1.00
22.51
C


ATOM
11024
CD
GLU
B
198
−72.635
−32.460
−23.522
1.00
25.57
C


ATOM
11025
OE1
GLU
B
198
−72.315
−31.896
−24.581
1.00
28.90
O


ATOM
11026
OE2
GLU
B
198
−73.326
−33.506
−23.540
1.00
28.09
O


ATOM
11027
C
GLU
B
198
−73.271
−29.101
−22.259
1.00
20.72
C


ATOM
11028
O
GLU
B
198
−74.281
−29.693
−22.644
1.00
20.82
O


ATOM
11030
N
ALA
B
199
−73.282
−28.199
−21.270
1.00
20.86
N


ATOM
11031
CA
ALA
B
199
−74.508
−27.814
−20.565
1.00
20.66
C


ATOM
11033
CB
ALA
B
199
−74.176
−27.013
−19.326
1.00
20.36
C


ATOM
11037
C
ALA
B
199
−75.472
−27.033
−21.461
1.00
20.84
C


ATOM
11038
O
ALA
B
199
−76.644
−27.404
−21.590
1.00
20.84
O


ATOM
11040
N
VAL
B
200
−75.002
−25.961
−22.092
1.00
21.12
N


ATOM
11041
CA
VAL
B
200
−75.903
−25.177
−22.942
1.00
21.60
C


ATOM
11043
CB
VAL
B
200
−75.215
−24.042
−23.701
1.00
21.30
C


ATOM
11045
CG1
VAL
B
200
−74.660
−23.055
−22.738
1.00
21.26
C


ATOM
11049
CG2
VAL
B
200
−74.135
−24.575
−24.626
1.00
21.28
C


ATOM
11053
C
VAL
B
200
−76.603
−26.077
−23.944
1.00
22.32
C


ATOM
11054
O
VAL
B
200
−77.791
−25.908
−24.218
1.00
22.32
O


ATOM
11056
N
TRP
B
201
−75.872
−27.047
−24.472
1.00
23.16
N


ATOM
11057
CA
TRP
B
201
−76.441
−27.946
−25.454
1.00
24.02
C


ATOM
11059
CB
TRP
B
201
−75.348
−28.718
−26.195
1.00
24.36
C


ATOM
11062
CG
TRP
B
201
−75.898
−29.510
−27.331
1.00
24.84
C


ATOM
11063
CD1
TRP
B
201
−76.060
−29.092
−28.613
1.00
25.41
C


ATOM
11065
NE1
TRP
B
201
−76.599
−30.094
−29.374
1.00
25.61
N


ATOM
11067
CE2
TRP
B
201
−76.807
−31.184
−28.579
1.00
25.34
C


ATOM
11068
CD2
TRP
B
201
−76.374
−30.847
−27.279
1.00
25.56
C


ATOM
11069
CE3
TRP
B
201
−76.466
−31.800
−26.264
1.00
27.13
C


ATOM
11071
CZ3
TRP
B
201
−76.989
−33.046
−26.574
1.00
28.66
C


ATOM
11073
CH2
TRP
B
201
−77.419
−33.347
−27.885
1.00
28.01
C


ATOM
11075
CZ2
TRP
B
201
−77.332
−32.425
−28.894
1.00
26.09
C


ATOM
11077
C
TRP
B
201
−77.411
−28.938
−24.833
1.00
24.22
C


ATOM
11078
O
TRP
B
201
−78.473
−29.172
−25.383
1.00
24.83
O


ATOM
11080
N
SER
B
202
−77.034
−29.541
−23.715
1.00
24.39
N


ATOM
11081
CA
SER
B
202
−77.818
−30.626
−23.144
1.00
24.50
C


ATOM
11083
CB
SER
B
202
−76.968
−31.455
−22.184
1.00
24.42
C


ATOM
11086
OG
SER
B
202
−75.848
−32.021
−22.848
1.00
23.80
O


ATOM
11088
C
SER
B
202
−79.067
−30.099
−22.441
1.00
24.98
C


ATOM
11089
O
SER
B
202
−80.057
−30.816
−22.324
1.00
24.73
O


ATOM
11091
N
ILE
B
203
−79.032
−28.849
−21.982
1.00
25.60
N


ATOM
11092
CA
ILE
B
203
−80.219
−28.255
−21.372
1.00
25.98
C


ATOM
11094
CB
ILE
B
203
−79.908
−26.967
−20.591
1.00
25.85
C


ATOM
11096
CG1
ILE
B
203
−79.086
−27.326
−19.348
1.00
25.76
C


ATOM
11099
CD1
ILE
B
203
−78.779
−26.159
−18.431
1.00
26.28
C


ATOM
11103
CG2
ILE
B
203
−81.212
−26.253
−20.206
1.00
24.51
C


ATOM
11107
C
ILE
B
203
−81.280
−28.016
−22.443
1.00
26.67
C


ATOM
11108
O
ILE
B
203
−82.428
−28.398
−22.260
1.00
26.96
O


ATOM
11110
N
GLU
B
204
−80.879
−27.412
−23.563
1.00
27.30
N


ATOM
11111
CA
GLU
B
204
−81.758
−27.205
−24.720
1.00
27.49
C


ATOM
11113
CB
GLU
B
204
−81.036
−26.384
−25.804
1.00
27.67
C


ATOM
11116
CG
GLU
B
204
−81.849
−26.077
−27.073
1.00
28.50
C


ATOM
11119
CD
GLU
B
204
−82.987
−25.072
−26.855
1.00
29.81
C


ATOM
11120
OE1
GLU
B
204
−83.158
−24.600
−25.711
1.00
31.40
O


ATOM
11121
OE2
GLU
B
204
−83.710
−24.750
−27.832
1.00
29.57
O


ATOM
11122
C
GLU
B
204
−82.201
−28.542
−25.287
1.00
27.52
C


ATOM
11123
O
GLU
B
204
−83.321
−28.667
−25.738
1.00
27.67
O


ATOM
11125
N
ALA
B
205
−81.326
−29.539
−25.266
1.00
27.74
N


ATOM
11126
CA
ALA
B
205
−81.684
−30.872
−25.728
1.00
28.09
C


ATOM
11128
CB
ALA
B
205
−80.472
−31.789
−25.726
1.00
27.75
C


ATOM
11132
C
ALA
B
205
−82.771
−31.448
−24.839
1.00
28.81
C


ATOM
11133
O
ALA
B
205
−83.811
−31.877
−25.325
1.00
28.90
O


ATOM
11135
N
TYR
B
206
−82.517
−31.433
−23.531
1.00
29.72
N


ATOM
11136
CA
TYR
B
206
−83.408
−32.020
−22.527
1.00
30.30
C


ATOM
11138
CB
TYR
B
206
−82.760
−31.915
−21.149
1.00
30.24
C


ATOM
11141
CG
TYR
B
206
−83.276
−32.889
−20.127
1.00
29.79
C


ATOM
11142
CD1
TYR
B
206
−83.055
−34.245
−20.286
1.00
30.71
C


ATOM
11144
CE1
TYR
B
206
−83.498
−35.161
−19.360
1.00
30.78
C


ATOM
11146
CZ
TYR
B
206
−84.159
−34.731
−18.238
1.00
30.03
C


ATOM
11147
OH
TYR
B
206
−84.582
−35.684
−17.343
1.00
30.51
O


ATOM
11149
CE2
TYR
B
206
−84.394
−33.379
−18.042
1.00
29.42
C


ATOM
11151
CD2
TYR
B
206
−83.945
−32.463
−18.989
1.00
29.05
C


ATOM
11153
C
TYR
B
206
−84.744
−31.308
−22.467
1.00
31.19
C


ATOM
11154
O
TYR
B
206
−85.790
−31.930
−22.311
1.00
31.44
O


ATOM
11156
N
ARG
B
207
−84.683
−29.988
−22.558
1.00
32.21
N


ATOM
11157
CA
ARG
B
207
−85.853
−29.128
−22.521
1.00
33.04
C


ATOM
11159
CB
ARG
B
207
−85.394
−27.700
−22.843
1.00
32.86
C


ATOM
11162
CG
ARG
B
207
−86.441
−26.637
−22.846
1.00
32.82
C


ATOM
11165
CD
ARG
B
207
−85.905
−25.360
−23.462
1.00
32.76
C


ATOM
11168
NE
ARG
B
207
−85.148
−24.561
−22.504
1.00
32.80
N


ATOM
11170
CZ
ARG
B
207
−85.676
−23.709
−21.628
1.00
33.40
C


ATOM
11171
NH1
ARG
B
207
−86.989
−23.526
−21.555
1.00
34.29
N


ATOM
11174
NH2
ARG
B
207
−84.884
−23.030
−20.808
1.00
33.89
N


ATOM
11177
C
ARG
B
207
−86.971
−29.606
−23.478
1.00
34.14
C


ATOM
11178
O
ARG
B
207
−88.144
−29.521
−23.128
1.00
34.34
O


ATOM
11180
N
LYS
B
208
−86.607
−30.137
−24.652
1.00
35.32
N


ATOM
11181
CA
LYS
B
208
−87.581
−30.535
−25.688
1.00
36.16
C


ATOM
11183
CB
LYS
B
208
−86.960
−30.468
−27.090
1.00
36.01
C


ATOM
11186
CG
LYS
B
208
−86.126
−29.234
−27.368
1.00
35.85
C


ATOM
11189
CD
LYS
B
208
−85.934
−29.023
−28.861
1.00
35.98
C


ATOM
11192
CE
LYS
B
208
−84.774
−28.086
−29.186
1.00
35.63
C


ATOM
11195
NZ
LYS
B
208
−83.566
−28.843
−29.596
1.00
35.17
N


ATOM
11199
C
LYS
B
208
−88.166
−31.937
−25.490
1.00
37.25
C


ATOM
11200
O
LYS
B
208
−89.212
−32.250
−26.054
1.00
37.50
O


ATOM
11202
N
LYS
B
209
−87.484
−32.788
−24.730
1.00
38.58
N


ATOM
11203
CA
LYS
B
209
−88.026
−34.106
−24.383
1.00
39.89
C


ATOM
11205
CB
LYS
B
209
−87.013
−34.923
−23.578
1.00
40.23
C


ATOM
11208
CG
LYS
B
209
−85.914
−35.604
−24.373
1.00
41.47
C


ATOM
11211
CD
LYS
B
209
−84.858
−36.178
−23.417
1.00
43.71
C


ATOM
11214
CE
LYS
B
209
−84.448
−37.612
−23.764
1.00
45.15
C


ATOM
11217
NZ
LYS
B
209
−85.390
−38.621
−23.163
1.00
45.73
N


ATOM
11221
C
LYS
B
209
−89.288
−33.950
−23.530
1.00
40.51
C


ATOM
11222
O
LYS
B
209
−89.230
−33.367
−22.441
1.00
40.84
O


ATOM
11224
N
GLU
B
210
−90.418
−34.477
−24.000
1.00
40.92
N


ATOM
11225
CA
GLU
B
210
−91.652
−34.386
−23.223
1.00
41.27
C


ATOM
11227
CB
GLU
B
210
−92.855
−34.935
−23.998
1.00
41.77
C


ATOM
11230
CG
GLU
B
210
−92.822
−36.446
−24.265
1.00
43.47
C


ATOM
11233
CD
GLU
B
210
−94.049
−36.922
−25.032
1.00
45.23
C


ATOM
11234
OE1
GLU
B
210
−95.188
−36.590
−24.612
1.00
45.02
O


ATOM
11235
OE2
GLU
B
210
−93.863
−37.629
−26.053
1.00
46.74
O


ATOM
11236
C
GLU
B
210
−91.494
−35.121
−21.895
1.00
40.71
C


ATOM
11237
O
GLU
B
210
−91.996
−34.667
−20.864
1.00
40.99
O


ATOM
11239
N
ASP
B
211
−90.773
−36.240
−21.924
1.00
39.76
N


ATOM
11240
CA
ASP
B
211
−90.533
−37.050
−20.721
1.00
38.96
C


ATOM
11242
CB
ASP
B
211
−90.151
−38.476
−21.123
1.00
39.10
C


ATOM
11245
CG
ASP
B
211
−89.101
−38.499
−22.212
1.00
40.26
C


ATOM
11246
OD1
ASP
B
211
−89.327
−37.827
−23.254
1.00
41.25
O


ATOM
11247
OD2
ASP
B
211
−88.055
−39.160
−22.020
1.00
41.66
O


ATOM
11248
C
ASP
B
211
−89.440
−36.468
−19.826
1.00
37.75
C


ATOM
11249
O
ASP
B
211
−89.009
−37.131
−18.892
1.00
37.62
O


ATOM
11251
N
ALA
B
212
−88.989
−35.245
−20.115
1.00
36.47
N


ATOM
11252
CA
ALA
B
212
−87.941
−34.587
−19.335
1.00
35.36
C


ATOM
11254
CB
ALA
B
212
−87.516
−33.294
−20.011
1.00
35.23
C


ATOM
11258
C
ALA
B
212
−88.423
−34.307
−17.920
1.00
34.40
C


ATOM
11259
O
ALA
B
212
−89.559
−33.870
−17.728
1.00
34.42
O


ATOM
11261
N
ASN
B
213
−87.565
−34.574
−16.937
1.00
33.24
N


ATOM
11262
CA
ASN
B
213
−87.890
−34.333
−15.533
1.00
32.67
C


ATOM
11264
CB
ASN
B
213
−86.840
−34.986
−14.623
1.00
32.58
C


ATOM
11267
CG
ASN
B
213
−87.204
−34.912
−13.152
1.00
32.67
C


ATOM
11268
OD1
ASN
B
213
−87.891
−33.994
−12.722
1.00
32.62
O


ATOM
11269
ND2
ASN
B
213
−86.734
−35.882
−12.371
1.00
33.18
N


ATOM
11272
C
ASN
B
213
−87.990
−32.825
−15.266
1.00
32.19
C


ATOM
11273
O
ASN
B
213
−87.010
−32.092
−15.376
1.00
32.38
O


ATOM
11275
N
GLN
B
214
−89.182
−32.356
−14.923
1.00
31.48
N


ATOM
11276
CA
GLN
B
214
−89.389
−30.924
−14.756
1.00
30.85
C


ATOM
11278
CB
GLN
B
214
−90.889
−30.581
−14.693
1.00
30.82
C


ATOM
11281
CG
GLN
B
214
−91.684
−30.869
−16.002
1.00
30.80
C


ATOM
11284
CD
GLN
B
214
−90.990
−30.378
−17.291
1.00
29.69
C


ATOM
11285
OE1
GLN
B
214
−90.893
−29.174
−17.543
1.00
29.07
O


ATOM
11286
NE2
GLN
B
214
−90.526
−31.321
−18.113
1.00
27.71
N


ATOM
11289
C
GLN
B
214
−88.638
−30.369
−13.543
1.00
30.37
C


ATOM
11290
O
GLN
B
214
−88.263
−29.200
−13.533
1.00
30.80
O


ATOM
11292
N
VAL
B
215
−88.393
−31.200
−12.532
1.00
29.54
N


ATOM
11293
CA
VAL
B
215
−87.566
−30.779
−11.395
1.00
28.56
C


ATOM
11295
CB
VAL
B
215
−87.564
−31.818
−10.260
1.00
28.64
C


ATOM
11297
CG1
VAL
B
215
−86.758
−31.299
−9.075
1.00
27.94
C


ATOM
11301
CG2
VAL
B
215
−88.999
−32.167
−9.856
1.00
28.26
C


ATOM
11305
C
VAL
B
215
−86.124
−30.545
−11.845
1.00
27.63
C


ATOM
11306
O
VAL
B
215
−85.566
−29.472
−11.615
1.00
27.58
O


ATOM
11308
N
LEU
B
216
−85.546
−31.548
−12.504
1.00
26.37
N


ATOM
11309
CA
LEU
B
216
−84.143
−31.509
−12.930
1.00
25.47
C


ATOM
11311
CB
LEU
B
216
−83.714
−32.866
−13.490
1.00
25.46
C


ATOM
11314
CG
LEU
B
216
−82.274
−32.995
−13.979
1.00
24.97
C


ATOM
11316
CD1
LEU
B
216
−81.289
−32.640
−12.883
1.00
24.62
C


ATOM
11320
CD2
LEU
B
216
−82.042
−34.407
−14.472
1.00
24.55
C


ATOM
11324
C
LEU
B
216
−83.861
−30.447
−13.972
1.00
24.68
C


ATOM
11325
O
LEU
B
216
−82.819
−29.820
−13.929
1.00
24.91
O


ATOM
11327
N
LEU
B
217
−84.776
−30.269
−14.916
1.00
23.79
N


ATOM
11328
CA
LEU
B
217
−84.625
−29.260
−15.960
1.00
23.13
C


ATOM
11330
CB
LEU
B
217
−85.765
−29.384
−16.972
1.00
23.36
C


ATOM
11333
CG
LEU
B
217
−85.808
−28.374
−18.123
1.00
23.18
C


ATOM
11335
CD1
LEU
B
217
−84.462
−28.268
−18.796
1.00
22.69
C


ATOM
11339
CD2
LEU
B
217
−86.861
−28.786
−19.126
1.00
23.38
C


ATOM
11343
C
LEU
B
217
−84.631
−27.853
−15.383
1.00
22.51
C


ATOM
11344
O
LEU
B
217
−83.903
−26.973
−15.855
1.00
22.08
O


ATOM
11346
N
GLU
B
218
−85.481
−27.647
−14.375
1.00
21.84
N


ATOM
11347
CA
GLU
B
218
−85.619
−26.339
−13.725
1.00
21.26
C


ATOM
11349
CB
GLU
B
218
−86.813
−26.341
−12.768
1.00
21.27
C


ATOM
11352
CG
GLU
B
218
−87.206
−24.956
−12.246
1.00
21.35
C


ATOM
11355
CD
GLU
B
218
−88.501
−24.964
−11.441
1.00
20.89
C


ATOM
11356
OE1
GLU
B
218
−89.097
−26.052
−11.263
1.00
20.47
O


ATOM
11357
OE2
GLU
B
218
−88.915
−23.874
−10.986
1.00
20.15
O


ATOM
11358
C
GLU
B
218
−84.348
−26.001
−12.969
1.00
20.58
C


ATOM
11359
O
GLU
B
218
−83.877
−24.865
−12.982
1.00
20.31
O


ATOM
11361
N
LEU
B
219
−83.802
−27.015
−12.312
1.00
19.87
N


ATOM
11362
CA
LEU
B
219
−82.559
−26.885
−11.592
1.00
19.12
C


ATOM
11364
CB
LEU
B
219
−82.310
−28.132
−10.746
1.00
18.58
C


ATOM
11367
CG
LEU
B
219
−81.159
−28.054
−9.762
1.00
17.07
C


ATOM
11369
CD1
LEU
B
219
−81.321
−26.847
−8.872
1.00
16.10
C


ATOM
11373
CD2
LEU
B
219
−81.107
−29.311
−8.954
1.00
15.22
C


ATOM
11377
C
LEU
B
219
−81.443
−26.675
−12.599
1.00
19.16
C


ATOM
11378
O
LEU
B
219
−80.684
−25.721
−12.478
1.00
19.42
O


ATOM
11380
N
ALA
B
220
−81.368
−27.546
−13.606
1.00
19.04
N


ATOM
11381
CA
ALA
B
220
−80.356
−27.431
−14.669
1.00
19.04
C


ATOM
11383
CB
ALA
B
220
−80.648
−28.398
−15.817
1.00
18.64
C


ATOM
11387
C
ALA
B
220
−80.257
−26.004
−15.196
1.00
19.06
C


ATOM
11388
O
ALA
B
220
−79.159
−25.472
−15.321
1.00
19.08
O


ATOM
11390
N
ILE
B
221
−81.410
−25.393
−15.475
1.00
19.19
N


ATOM
11391
CA
ILE
B
221
−81.473
−24.016
−15.963
1.00
19.22
C


ATOM
11393
CB
ILE
B
221
−82.898
−23.638
−16.423
1.00
19.03
C


ATOM
11395
CG1
ILE
B
221
−83.229
−24.323
−17.755
1.00
18.52
C


ATOM
11398
CD1
ILE
B
221
−84.687
−24.645
−17.928
1.00
17.52
C


ATOM
11402
CG2
ILE
B
221
−83.024
−22.121
−16.564
1.00
18.03
C


ATOM
11406
C
ILE
B
221
−81.030
−23.013
−14.906
1.00
19.62
C


ATOM
11407
O
ILE
B
221
−80.137
−22.190
−15.136
1.00
19.53
O


ATOM
11409
N
LEU
B
222
−81.666
−23.096
−13.745
1.00
20.27
N


ATOM
11410
CA
LEU
B
222
−81.412
−22.161
−12.654
1.00
20.63
C


ATOM
11412
CB
LEU
B
222
−82.203
−22.561
−11.412
1.00
20.50
C


ATOM
11415
CG
LEU
B
222
−82.195
−21.523
−10.296
1.00
20.76
C


ATOM
11417
CD1
LEU
B
222
−83.439
−21.675
−9.431
1.00
21.02
C


ATOM
11421
CD2
LEU
B
222
−80.924
−21.625
−9.453
1.00
20.21
C


ATOM
11425
C
LEU
B
222
−79.925
−22.089
−12.341
1.00
20.98
C


ATOM
11426
O
LEU
B
222
−79.337
−21.011
−12.381
1.00
21.40
O


ATOM
11428
N
ASP
B
223
−79.319
−23.240
−12.064
1.00
21.24
N


ATOM
11429
CA
ASP
B
223
−77.907
−23.298
−11.709
1.00
21.48
C


ATOM
11431
CB
ASP
B
223
−77.509
−24.724
−11.314
1.00
21.66
C


ATOM
11434
CG
ASP
B
223
−76.168
−24.792
−10.589
1.00
22.70
C


ATOM
11435
OD1
ASP
B
223
−75.095
−24.775
−11.244
1.00
24.25
O


ATOM
11436
OD2
ASP
B
223
−76.190
−24.893
−9.349
1.00
24.85
O


ATOM
11437
C
ASP
B
223
−77.017
−22.768
−12.835
1.00
21.64
C


ATOM
11438
O
ASP
B
223
−76.035
−22.104
−12.548
1.00
21.66
O


ATOM
11440
N
TYR
B
224
−77.361
−23.023
−14.100
1.00
22.07
N


ATOM
11441
CA
TYR
B
224
−76.517
−22.561
−15.218
1.00
22.47
C


ATOM
11443
CB
TYR
B
224
−76.980
−23.111
−16.589
1.00
22.35
C


ATOM
11446
CG
TYR
B
224
−76.032
−22.733
−17.724
1.00
22.25
C


ATOM
11447
CD1
TYR
B
224
−75.002
−23.576
−18.116
1.00
21.46
C


ATOM
11449
CE1
TYR
B
224
−74.126
−23.216
−19.135
1.00
21.43
C


ATOM
11451
CZ
TYR
B
224
−74.260
−21.991
−19.761
1.00
21.62
C


ATOM
11452
OH
TYR
B
224
−73.387
−21.618
−20.760
1.00
20.46
O


ATOM
11454
CE2
TYR
B
224
−75.270
−21.132
−19.387
1.00
22.04
C


ATOM
11456
CD2
TYR
B
224
−76.144
−21.500
−18.373
1.00
22.73
C


ATOM
11458
C
TYR
B
224
−76.414
−21.023
−15.262
1.00
23.06
C


ATOM
11459
O
TYR
B
224
−75.323
−20.467
−15.507
1.00
22.89
O


ATOM
11461
N
ASN
B
225
−77.543
−20.350
−15.027
1.00
23.66
N


ATOM
11462
CA
ASN
B
225
−77.593
−18.884
−15.072
1.00
24.16
C


ATOM
11464
CB
ASN
B
225
−79.040
−18.367
−15.107
1.00
24.12
C


ATOM
11467
CG
ASN
B
225
−79.778
−18.761
−16.365
1.00
23.89
C


ATOM
11468
OD1
ASN
B
225
−79.182
−18.877
−17.437
1.00
24.33
O


ATOM
11469
ND2
ASN
B
225
−81.088
−18.965
−16.244
1.00
22.05
N


ATOM
11472
C
ASN
B
225
−76.880
−18.265
−13.881
1.00
24.73
C


ATOM
11473
O
ASN
B
225
−76.195
−17.247
−14.027
1.00
25.09
O


ATOM
11475
N
MET
B
226
−77.067
−18.864
−12.703
1.00
25.15
N


ATOM
11476
CA
MET
B
226
−76.423
−18.390
−11.479
1.00
25.53
C


ATOM
11478
CB
MET
B
226
−76.806
−19.278
−10.282
1.00
26.11
C


ATOM
11481
CG
MET
B
226
−75.905
−19.169
−9.023
1.00
27.59
C


ATOM
11484
SD
MET
B
226
−75.676
−20.788
−8.214
1.00
30.67
S


ATOM
11485
CE
MET
B
226
−77.308
−21.005
−7.476
1.00
29.78
C


ATOM
11489
C
MET
B
226
−74.920
−18.380
−11.692
1.00
25.16
C


ATOM
11490
O
MET
B
226
−74.258
−17.387
−11.391
1.00
25.10
O


ATOM
11492
N
ILE
B
227
−74.388
−19.473
−12.239
1.00
24.88
N


ATOM
11493
CA
ILE
B
227
−72.944
−19.564
−12.501
1.00
24.70
C


ATOM
11495
CB
ILE
B
227
−72.476
−20.994
−12.882
1.00
24.45
C


ATOM
11497
CG1
ILE
B
227
−72.656
−21.947
−11.695
1.00
23.80
C


ATOM
11500
CD1
ILE
B
227
−72.094
−23.332
−11.885
1.00
21.97
C


ATOM
11504
CG2
ILE
B
227
−71.030
−20.969
−13.266
1.00
24.90
C


ATOM
11508
C
ILE
B
227
−72.529
−18.550
−13.566
1.00
24.45
C


ATOM
11509
O
ILE
B
227
−71.578
−17.810
−13.373
1.00
24.42
O


ATOM
11511
N
GLN
B
228
−73.264
−18.484
−14.668
1.00
24.31
N


ATOM
11512
CA
GLN
B
228
−73.038
−17.414
−15.639
1.00
24.19
C


ATOM
11514
CB
GLN
B
228
−74.143
−17.363
−16.688
1.00
24.02
C


ATOM
11517
CG
GLN
B
228
−73.788
−16.488
−17.864
1.00
23.42
C


ATOM
11520
CD
GLN
B
228
−74.807
−16.567
−18.983
1.00
23.70
C


ATOM
11521
OE1
GLN
B
228
−75.790
−15.813
−19.015
1.00
24.07
O


ATOM
11522
NE2
GLN
B
228
−74.564
−17.465
−19.927
1.00
23.49
N


ATOM
11525
C
GLN
B
228
−72.913
−16.037
−14.987
1.00
24.37
C


ATOM
11526
O
GLN
B
228
−72.145
−15.210
−15.470
1.00
24.45
O


ATOM
11528
N
SER
B
229
−73.653
−15.776
−13.909
1.00
24.45
N


ATOM
11529
CA
SER
B
229
−73.578
−14.459
−13.264
1.00
24.76
C


ATOM
11531
CB
SER
B
229
−74.815
−14.163
−12.397
1.00
24.84
C


ATOM
11534
OG
SER
B
229
−74.711
−14.718
−11.096
1.00
25.81
O


ATOM
11536
C
SER
B
229
−72.270
−14.279
−12.469
1.00
24.64
C


ATOM
11537
O
SER
B
229
−71.726
−13.172
−12.408
1.00
24.95
O


ATOM
11539
N
VAL
B
230
−71.751
−15.350
−11.878
1.00
24.20
N


ATOM
11540
CA
VAL
B
230
−70.415
−15.277
−11.299
1.00
23.99
C


ATOM
11542
CB
VAL
B
230
−70.006
−16.581
−10.574
1.00
23.92
C


ATOM
11544
CG1
VAL
B
230
−68.546
−16.510
−10.105
1.00
23.71
C


ATOM
11548
CG2
VAL
B
230
−70.923
−16.853
−9.409
1.00
23.52
C


ATOM
11552
C
VAL
B
230
−69.386
−14.966
−12.400
1.00
24.12
C


ATOM
11553
O
VAL
B
230
−68.397
−14.291
−12.136
1.00
24.01
O


ATOM
11555
N
TYR
B
231
−69.612
−15.461
−13.621
1.00
24.30
N


ATOM
11556
CA
TYR
B
231
−68.672
−15.235
−14.728
1.00
24.53
C


ATOM
11558
CB
TYR
B
231
−68.997
−16.104
−15.946
1.00
24.08
C


ATOM
11561
CG
TYR
B
231
−68.892
−17.605
−15.754
1.00
23.29
C


ATOM
11562
CD1
TYR
B
231
−68.173
−18.168
−14.706
1.00
23.08
C


ATOM
11564
CE1
TYR
B
231
−68.073
−19.547
−14.565
1.00
22.01
C


ATOM
11566
CZ
TYR
B
231
−68.685
−20.369
−15.479
1.00
21.16
C


ATOM
11567
OH
TYR
B
231
−68.607
−21.746
−15.353
1.00
20.18
O


ATOM
11569
CE2
TYR
B
231
−69.387
−19.825
−16.523
1.00
21.64
C


ATOM
11571
CD2
TYR
B
231
−69.476
−18.464
−16.662
1.00
22.15
C


ATOM
11573
C
TYR
B
231
−68.658
−13.772
−15.166
1.00
25.37
C


ATOM
11574
O
TYR
B
231
−67.602
−13.221
−15.528
1.00
25.25
O


ATOM
11576
N
GLN
B
232
−69.832
−13.148
−15.129
1.00
26.11
N


ATOM
11577
CA
GLN
B
232
−69.978
−11.797
−15.622
1.00
26.68
C


ATOM
11579
CB
GLN
B
232
−71.430
−11.541
−15.995
1.00
26.45
C


ATOM
11582
CG
GLN
B
232
−71.883
−12.385
−17.176
1.00
25.67
C


ATOM
11585
CD
GLN
B
232
−73.388
−12.394
−17.381
1.00
24.91
C


ATOM
11586
OE1
GLN
B
232
−74.140
−11.761
−16.637
1.00
24.81
O


ATOM
11587
NE2
GLN
B
232
−73.834
−13.113
−18.407
1.00
23.39
N


ATOM
11590
C
GLN
B
232
−69.445
−10.802
−14.593
1.00
27.90
C


ATOM
11591
O
GLN
B
232
−68.909
−9.752
−14.958
1.00
27.96
O


ATOM
11593
N
ARG
B
233
−69.572
−11.145
−13.312
1.00
29.35
N


ATOM
11594
CA
ARG
B
233
−68.908
−10.396
−12.241
1.00
30.59
C


ATOM
11596
CB
ARG
B
233
−69.490
−10.775
−10.867
1.00
30.99
C


ATOM
11599
CG
ARG
B
233
−68.824
−10.114
−9.661
1.00
32.72
C


ATOM
11602
CD
ARG
B
233
−69.695
−10.210
−8.391
1.00
35.70
C


ATOM
11605
NE
ARG
B
233
−70.162
−11.578
−8.086
1.00
38.29
N


ATOM
11607
CZ
ARG
B
233
−71.403
−12.053
−8.276
1.00
39.65
C


ATOM
11608
NH1
ARG
B
233
−72.367
−11.287
−8.787
1.00
40.23
N


ATOM
11611
NH2
ARG
B
233
−71.687
−13.320
−7.949
1.00
39.61
N


ATOM
11614
C
ARG
B
233
−67.390
−10.632
−12.303
1.00
31.10
C


ATOM
11615
O
ARG
B
233
−66.615
−9.693
−12.181
1.00
31.19
O


ATOM
11617
N
ASP
B
234
−66.954
−11.868
−12.517
1.00
31.91
N


ATOM
11618
CA
ASP
B
234
−65.530
−12.102
−12.750
1.00
32.63
C


ATOM
11620
CB
ASP
B
234
−65.220
−13.566
−13.090
1.00
32.53
C


ATOM
11623
CG
ASP
B
234
−65.336
−14.493
−11.894
1.00
32.90
C


ATOM
11624
OD1
ASP
B
234
−65.477
−14.028
−10.746
1.00
33.06
O


ATOM
11625
OD2
ASP
B
234
−65.293
−15.713
−12.107
1.00
34.22
O


ATOM
11626
C
ASP
B
234
−65.058
−11.206
−13.886
1.00
33.10
C


ATOM
11627
O
ASP
B
234
−64.097
−10.458
−13.723
1.00
33.55
O


ATOM
11629
N
LEU
B
235
−65.753
−11.256
−15.017
1.00
33.49
N


ATOM
11630
CA
LEU
B
235
−65.311
−10.550
−16.216
1.00
34.06
C


ATOM
11632
CB
LEU
B
235
−66.139
−10.996
−17.422
1.00
33.80
C


ATOM
11635
CG
LEU
B
235
−65.769
−10.403
−18.775
1.00
32.13
C


ATOM
11637
CD1
LEU
B
235
−64.303
−10.679
−19.050
1.00
30.94
C


ATOM
11641
CD2
LEU
B
235
−66.670
−10.959
−19.870
1.00
29.93
C


ATOM
11645
C
LEU
B
235
−65.345
−9.023
−16.103
1.00
35.40
C


ATOM
11646
O
LEU
B
235
−64.500
−8.352
−16.682
1.00
35.61
O


ATOM
11648
N
ARG
B
236
−66.327
−8.473
−15.386
1.00
36.92
N


ATOM
11649
CA
ARG
B
236
−66.380
−7.019
−15.137
1.00
38.06
C


ATOM
11651
CB
ARG
B
236
−67.650
−6.607
−14.367
1.00
38.43
C


ATOM
11654
CG
ARG
B
236
−68.882
−6.321
−15.230
1.00
39.58
C


ATOM
11657
CD
ARG
B
236
−69.958
−5.520
−14.459
1.00
40.93
C


ATOM
11660
NE
ARG
B
236
−70.383
−6.149
−13.199
1.00
41.74
N


ATOM
11662
CZ
ARG
B
236
−71.208
−7.195
−13.095
1.00
42.20
C


ATOM
11663
NH1
ARG
B
236
−71.719
−7.784
−14.175
1.00
41.61
N


ATOM
11666
NH2
ARG
B
236
−71.517
−7.672
−11.892
1.00
42.99
N


ATOM
11669
C
ARG
B
236
−65.170
−6.550
−14.343
1.00
38.57
C


ATOM
11670
O
ARG
B
236
−64.593
−5.524
−14.665
1.00
38.61
O


ATOM
11672
N
GLU
B
237
−64.822
−7.292
−13.291
1.00
39.42
N


ATOM
11673
CA
GLU
B
237
−63.625
−7.017
−12.480
1.00
40.18
C


ATOM
11675
CB
GLU
B
237
−63.471
−8.033
−11.317
1.00
40.67
C


ATOM
11678
CG
GLU
B
237
−64.336
−7.750
−10.057
1.00
42.61
C


ATOM
11681
CD
GLU
B
237
−64.258
−8.858
−8.961
1.00
45.07
C


ATOM
11682
OE1
GLU
B
237
−64.051
−10.066
−9.282
1.00
46.74
O


ATOM
11683
OE2
GLU
B
237
−64.436
−8.511
−7.764
1.00
45.87
O


ATOM
11684
C
GLU
B
237
−62.352
−7.024
−13.344
1.00
39.93
C


ATOM
11685
O
GLU
B
237
−61.593
−6.050
−13.332
1.00
39.96
O


ATOM
11687
N
THR
B
238
−62.123
−8.104
−14.095
1.00
39.56
N


ATOM
11688
CA
THR
B
238
−60.900
−8.200
−14.895
1.00
39.42
C


ATOM
11690
CB
THR
B
238
−60.492
−9.666
−15.268
1.00
39.39
C


ATOM
11692
OG1
THR
B
238
−61.222
−10.123
−16.413
1.00
39.37
O


ATOM
11694
CG2
THR
B
238
−60.683
−10.612
−14.095
1.00
38.73
C


ATOM
11698
C
THR
B
238
−60.961
−7.326
−16.156
1.00
39.48
C


ATOM
11699
O
THR
B
238
−59.930
−7.062
−16.761
1.00
39.48
O


ATOM
11701
N
SER
B
239
−62.149
−6.871
−16.549
1.00
39.62
N


ATOM
11702
CA
SER
B
239
−62.264
−5.882
−17.631
1.00
39.63
C


ATOM
11704
CB
SER
B
239
−63.689
−5.810
−18.172
1.00
39.56
C


ATOM
11707
OG
SER
B
239
−63.945
−6.917
−19.010
1.00
38.96
O


ATOM
11709
C
SER
B
239
−61.796
−4.496
−17.181
1.00
39.99
C


ATOM
11710
O
SER
B
239
−61.108
−3.807
−17.933
1.00
40.10
O


ATOM
11712
N
ARG
B
240
−62.168
−4.088
−15.965
1.00
40.35
N


ATOM
11713
CA
ARG
B
240
−61.624
−2.864
−15.363
1.00
40.68
C


ATOM
11715
CB
ARG
B
240
−62.025
−2.712
−13.881
1.00
41.26
C


ATOM
11718
CG
ARG
B
240
−63.231
−1.782
−13.621
1.00
43.86
C


ATOM
11721
CD
ARG
B
240
−63.334
−1.375
−12.130
1.00
47.07
C


ATOM
11724
NE
ARG
B
240
−63.365
−2.537
−11.222
1.00
50.14
N


ATOM
11726
CZ
ARG
B
240
−64.462
−3.077
−10.671
1.00
52.32
C


ATOM
11727
NH1
ARG
B
240
−64.342
−4.141
−9.875
1.00
53.16
N


ATOM
11730
NH2
ARG
B
240
−65.677
−2.577
−10.899
1.00
53.18
N


ATOM
11733
C
ARG
B
240
−60.113
−2.914
−15.464
1.00
39.91
C


ATOM
11734
O
ARG
B
240
−59.490
−2.000
−15.988
1.00
39.86
O


ATOM
11736
N
TRP
B
241
−59.541
−4.002
−14.962
1.00
39.14
N


ATOM
11737
CA
TRP
B
241
−58.105
−4.217
−15.004
1.00
38.37
C


ATOM
11739
CB
TRP
B
241
−57.773
−5.635
−14.521
1.00
38.17
C


ATOM
11742
CG
TRP
B
241
−56.373
−6.005
−14.773
1.00
36.58
C


ATOM
11743
CD1
TRP
B
241
−55.290
−5.668
−14.022
1.00
36.32
C


ATOM
11745
NE1
TRP
B
241
−54.152
−6.176
−14.587
1.00
35.63
N


ATOM
11747
CE2
TRP
B
241
−54.494
−6.846
−15.731
1.00
34.31
C


ATOM
11748
CD2
TRP
B
241
−55.883
−6.757
−15.874
1.00
33.99
C


ATOM
11749
CE3
TRP
B
241
−56.484
−7.357
−16.977
1.00
33.28
C


ATOM
11751
CZ3
TRP
B
241
−55.691
−8.034
−17.883
1.00
32.79
C


ATOM
11753
CH2
TRP
B
241
−54.313
−8.101
−17.720
1.00
33.06
C


ATOM
11755
CZ2
TRP
B
241
−53.697
−7.515
−16.648
1.00
34.16
C


ATOM
11757
C
TRP
B
241
−57.561
−3.999
−16.409
1.00
38.25
C


ATOM
11758
O
TRP
B
241
−56.607
−3.252
−16.603
1.00
38.13
O


ATOM
11760
N
TRP
B
242
−58.186
−4.644
−17.386
1.00
38.21
N


ATOM
11761
CA
TRP
B
242
−57.695
−4.628
−18.765
1.00
38.27
C


ATOM
11763
CB
TRP
B
242
−58.479
−5.640
−19.609
1.00
38.31
C


ATOM
11766
CG
TRP
B
242
−57.948
−5.872
−20.990
1.00
38.61
C


ATOM
11767
CD1
TRP
B
242
−58.642
−5.754
−22.156
1.00
39.11
C


ATOM
11769
NE1
TRP
B
242
−57.828
−6.046
−23.224
1.00
39.31
N


ATOM
11771
CE2
TRP
B
242
−56.582
−6.363
−22.759
1.00
38.95
C


ATOM
11772
CD2
TRP
B
242
−56.617
−6.263
−21.356
1.00
38.66
C


ATOM
11773
CE3
TRP
B
242
−55.454
−6.528
−20.637
1.00
38.98
C


ATOM
11775
CZ3
TRP
B
242
−54.314
−6.886
−21.326
1.00
39.20
C


ATOM
11777
CH2
TRP
B
242
−54.313
−6.985
−22.716
1.00
39.12
C


ATOM
11779
CZ2
TRP
B
242
−55.434
−6.724
−23.451
1.00
39.22
C


ATOM
11781
C
TRP
B
242
−57.739
−3.241
−19.400
1.00
38.27
C


ATOM
11782
O
TRP
B
242
−56.814
−2.873
−20.108
1.00
37.81
O


ATOM
11784
N
ARG
B
243
−58.809
−2.489
−19.140
1.00
38.79
N


ATOM
11785
CA
ARG
B
243
−58.936
−1.097
−19.607
1.00
39.37
C


ATOM
11787
CB
ARG
B
243
−60.351
−.552
−19.346
1.00
39.66
C


ATOM
11790
CG
ARG
B
243
−61.324
−.792
−20.520
1.00
41.96
C


ATOM
11793
CD
ARG
B
243
−62.687
−1.338
−20.085
1.00
44.82
C


ATOM
11796
NE
ARG
B
243
−63.461
−.380
−19.296
1.00
47.15
N


ATOM
11798
CZ
ARG
B
243
−64.389
−.704
−18.386
1.00
49.26
C


ATOM
11799
NH1
ARG
B
243
−64.685
−1.976
−18.109
1.00
49.18
N


ATOM
11802
NH2
ARG
B
243
−65.028
.262
−17.729
1.00
50.40
N


ATOM
11805
C
ARG
B
243
−57.890
−.183
−18.980
1.00
39.28
C


ATOM
11806
O
ARG
B
243
−57.246
.577
−19.681
1.00
39.40
O


ATOM
11808
N
ARG
B
244
−57.724
−.282
−17.665
1.00
39.52
N


ATOM
11809
CA
ARG
B
244
−56.734
.489
−16.888
1.00
39.61
C


ATOM
11811
CB
ARG
B
244
−56.774
.041
−15.409
1.00
40.25
C


ATOM
11814
CG
ARG
B
244
−56.115
.971
−14.383
1.00
42.01
C


ATOM
11817
CD
ARG
B
244
−57.029
2.154
−14.017
1.00
44.39
C


ATOM
11820
NE
ARG
B
244
−56.391
3.115
−13.103
1.00
46.36
N


ATOM
11822
CZ
ARG
B
244
−55.434
3.984
−13.441
1.00
48.06
C


ATOM
11823
NH1
ARG
B
244
−54.951
4.043
−14.686
1.00
48.90
N


ATOM
11826
NH2
ARG
B
244
−54.947
4.805
−12.519
1.00
48.82
N


ATOM
11829
C
ARG
B
244
−55.323
.322
−17.437
1.00
38.83
C


ATOM
11830
O
ARG
B
244
−54.594
1.300
−17.567
1.00
38.39
O


ATOM
11832
N
VAL
B
245
−54.953
−.922
−17.738
1.00
38.52
N


ATOM
11833
CA
VAL
B
245
−53.694
−1.241
−18.437
1.00
38.43
C


ATOM
11835
CB
VAL
B
245
−53.491
−2.760
−18.614
1.00
38.31
C


ATOM
11837
CG1
VAL
B
245
−53.247
−3.426
−17.283
1.00
37.77
C


ATOM
11841
CG2
VAL
B
245
−52.344
−3.036
−19.561
1.00
37.66
C


ATOM
11845
C
VAL
B
245
−53.650
−.638
−19.832
1.00
38.68
C


ATOM
11846
O
VAL
B
245
−52.615
−.153
−20.247
1.00
38.57
O


ATOM
11848
N
GLY
B
246
−54.765
−.724
−20.559
1.00
39.26
N


ATOM
11849
CA
GLY
B
246
−54.974
−.012
−21.834
1.00
39.62
C


ATOM
11852
C
GLY
B
246
−53.962
−.262
−22.942
1.00
40.08
C


ATOM
11853
O
GLY
B
246
−53.617
.650
−23.700
1.00
39.89
O


ATOM
11855
N
LEU
B
247
−53.502
−1.500
−23.062
1.00
40.75
N


ATOM
11856
CA
LEU
B
247
−52.347
−1.778
−23.905
1.00
41.34
C


ATOM
11858
CB
LEU
B
247
−51.655
−3.065
−23.459
1.00
41.15
C


ATOM
11861
CG
LEU
B
247
−50.132
−3.026
−23.298
1.00
40.49
C


ATOM
11863
CD1
LEU
B
247
−49.622
−1.769
−22.605
1.00
39.50
C


ATOM
11867
CD2
LEU
B
247
−49.695
−4.242
−22.526
1.00
39.98
C


ATOM
11871
C
LEU
B
247
−52.744
−1.836
−25.370
1.00
42.50
C


ATOM
11872
O
LEU
B
247
−52.005
−1.358
−26.225
1.00
42.33
O


ATOM
11874
N
ALA
B
248
−53.925
−2.390
−25.654
1.00
44.14
N


ATOM
11875
CA
ALA
B
248
−54.439
−2.494
−27.041
1.00
45.16
C


ATOM
11877
CB
ALA
B
248
−55.705
−3.347
−27.087
1.00
45.15
C


ATOM
11881
C
ALA
B
248
−54.692
−1.140
−27.739
1.00
45.98
C


ATOM
11882
O
ALA
B
248
−54.604
−1.053
−28.971
1.00
46.37
O


ATOM
11884
N
THR
B
249
−55.004
−.095
−26.975
1.00
46.64
N


ATOM
11885
CA
THR
B
249
−55.142
1.234
−27.570
1.00
47.25
C


ATOM
11887
CB
THR
B
249
−55.905
2.253
−26.655
1.00
47.44
C


ATOM
11889
OG1
THR
B
249
−55.001
2.845
−25.706
1.00
47.52
O


ATOM
11891
CG2
THR
B
249
−57.104
1.592
−25.928
1.00
47.44
C


ATOM
11895
C
THR
B
249
−53.756
1.793
−27.931
1.00
47.59
C


ATOM
11896
O
THR
B
249
−53.553
2.279
−29.049
1.00
48.07
O


ATOM
11898
N
LYS
B
250
−52.808
1.710
−26.995
1.00
47.67
N


ATOM
11899
CA
LYS
B
250
−51.469
2.299
−27.185
1.00
47.68
C


ATOM
11901
CB
LYS
B
250
−50.793
2.553
−25.833
1.00
47.68
C


ATOM
11904
CG
LYS
B
250
−51.428
3.673
−24.999
1.00
47.77
C


ATOM
11907
CD
LYS
B
250
−51.142
5.086
−25.552
1.00
47.59
C


ATOM
11910
CE
LYS
B
250
−49.676
5.496
−25.441
1.00
46.75
C


ATOM
11913
NZ
LYS
B
250
−49.479
6.878
−25.948
1.00
46.49
N


ATOM
11917
C
LYS
B
250
−50.541
1.465
−28.082
1.00
47.72
C


ATOM
11918
O
LYS
B
250
−49.591
1.988
−28.663
1.00
47.54
O


ATOM
11920
N
LEU
B
251
−50.804
.169
−28.184
1.00
47.82
N


ATOM
11921
CA
LEU
B
251
−50.072
−.675
−29.118
1.00
47.96
C


ATOM
11923
CB
LEU
B
251
−49.584
−1.970
−28.447
1.00
48.04
C


ATOM
11926
CG
LEU
B
251
−48.109
−2.064
−28.033
1.00
47.39
C


ATOM
11928
CD1
LEU
B
251
−47.659
−.881
−27.186
1.00
46.47
C


ATOM
11932
CD2
LEU
B
251
−47.894
−3.376
−27.304
1.00
46.78
C


ATOM
11936
C
LEU
B
251
−50.985
−.964
−30.306
1.00
48.19
C


ATOM
11937
O
LEU
B
251
−51.975
−1.693
−30.194
1.00
48.38
O


ATOM
11939
N
HIS
B
252
−50.627
−.396
−31.449
1.00
48.42
N


ATOM
11940
CA
HIS
B
252
−51.521
−.329
−32.601
1.00
48.62
C


ATOM
11942
CB
HIS
B
252
−51.110
.850
−33.492
1.00
48.99
C


ATOM
11945
CG
HIS
B
252
−51.000
2.145
−32.742
1.00
50.56
C


ATOM
11946
ND1
HIS
B
252
−52.095
2.935
−32.453
1.00
52.11
N


ATOM
11948
CE1
HIS
B
252
−51.704
3.993
−31.766
1.00
52.68
C


ATOM
11950
NE2
HIS
B
252
−50.397
3.912
−31.585
1.00
52.86
N


ATOM
11952
CD2
HIS
B
252
−49.932
2.764
−32.181
1.00
51.68
C


ATOM
11954
C
HIS
B
252
−51.582
−1.636
−33.383
1.00
48.14
C


ATOM
11955
O
HIS
B
252
−52.615
−1.975
−33.935
1.00
48.02
O


ATOM
11957
N
PHE
B
253
−50.481
−2.372
−33.405
1.00
48.10
N


ATOM
11958
CA
PHE
B
253
−50.422
−3.702
−34.044
1.00
48.18
C


ATOM
11960
CB
PHE
B
253
−48.965
−4.070
−34.375
1.00
48.16
C


ATOM
11963
CG
PHE
B
253
−48.119
−4.311
−33.163
1.00
47.85
C


ATOM
11964
CD1
PHE
B
253
−47.900
−5.596
−32.702
1.00
48.56
C


ATOM
11966
CE1
PHE
B
253
−47.140
−5.812
−31.576
1.00
48.98
C


ATOM
11968
CZ
PHE
B
253
−46.602
−4.729
−30.895
1.00
48.50
C


ATOM
11970
CE2
PHE
B
253
−46.817
−3.454
−31.349
1.00
47.40
C


ATOM
11972
CD2
PHE
B
253
−47.569
−3.249
−32.465
1.00
47.22
C


ATOM
11974
C
PHE
B
253
−51.038
−4.819
−33.185
1.00
48.30
C


ATOM
11975
O
PHE
B
253
−51.344
−5.903
−33.691
1.00
47.63
O


ATOM
11977
N
ALA
B
254
−51.204
−4.541
−31.888
1.00
48.86
N


ATOM
11978
CA
ALA
B
254
−51.653
−5.531
−30.903
1.00
49.13
C


ATOM
11980
CB
ALA
B
254
−51.290
−5.071
−29.494
1.00
49.02
C


ATOM
11984
C
ALA
B
254
−53.153
−5.806
−30.982
1.00
49.46
C


ATOM
11985
O
ALA
B
254
−53.967
−4.870
−31.011
1.00
49.50
O


ATOM
11987
N
ARG
B
255
−53.501
−7.097
−31.006
1.00
49.75
N


ATOM
11988
CA
ARG
B
255
−54.896
−7.549
−30.918
1.00
49.89
C


ATOM
11990
CB
ARG
B
255
−55.028
−9.051
−31.245
1.00
49.94
C


ATOM
11993
CG
ARG
B
255
−54.839
−9.439
−32.724
1.00
49.93
C


ATOM
11996
CD
ARG
B
255
−54.709
−10.967
−32.904
1.00
49.79
C


ATOM
11999
NE
ARG
B
255
−53.527
−11.505
−32.219
1.00
50.11
N


ATOM
12001
CZ
ARG
B
255
−53.276
−12.801
−32.010
1.00
50.54
C


ATOM
12002
NH1
ARG
B
255
−54.117
−13.736
−32.439
1.00
51.36
N


ATOM
12005
NH2
ARG
B
255
−52.175
−13.172
−31.360
1.00
50.21
N


ATOM
12008
C
ARG
B
255
−55.425
−7.308
−29.511
1.00
49.83
C


ATOM
12009
O
ARG
B
255
−54.666
−6.975
−28.599
1.00
49.95
O


ATOM
12011
N
ASP
B
256
−56.736
−7.463
−29.350
1.00
49.72
N


ATOM
12012
CA
ASP
B
256
−57.360
−7.483
−28.034
1.00
49.55
C


ATOM
12014
CB
ASP
B
256
−58.183
−6.222
−27.812
1.00
49.52
C


ATOM
12017
CG
ASP
B
256
−59.134
−6.362
−26.654
1.00
50.30
C


ATOM
12018
OD1
ASP
B
256
−60.310
−5.944
−26.780
1.00
50.45
O


ATOM
12019
OD2
ASP
B
256
−58.702
−6.926
−25.622
1.00
51.68
O


ATOM
12020
C
ASP
B
256
−58.254
−8.721
−27.931
1.00
49.15
C


ATOM
12021
O
ASP
B
256
−59.159
−8.901
−28.737
1.00
49.28
O


ATOM
12023
N
ARG
B
257
−57.998
−9.565
−26.937
1.00
48.61
N


ATOM
12024
CA
ARG
B
257
−58.743
−10.809
−26.765
1.00
48.20
C


ATOM
12026
CB
ARG
B
257
−57.949
−11.984
−27.373
1.00
48.41
C


ATOM
12029
CG
ARG
B
257
−57.505
−11.799
−28.821
1.00
49.37
C


ATOM
12032
CD
ARG
B
257
−58.693
−11.636
−29.784
1.00
50.80
C


ATOM
12035
NE
ARG
B
257
−58.908
−12.814
−30.627
1.00
52.11
N


ATOM
12037
CZ
ARG
B
257
−58.678
−12.876
−31.944
1.00
52.99
C


ATOM
12038
NH1
ARG
B
257
−58.214
−11.820
−32.623
1.00
52.37
N


ATOM
12041
NH2
ARG
B
257
−58.918
−14.016
−32.596
1.00
52.97
N


ATOM
12044
C
ARG
B
257
−59.032
−11.089
−25.280
1.00
47.32
C


ATOM
12045
O
ARG
B
257
−58.579
−12.101
−24.735
1.00
47.46
O


ATOM
12047
N
LEU
B
258
−59.770
−10.201
−24.616
1.00
45.94
N


ATOM
12048
CA
LEU
B
258
−60.108
−10.430
−23.205
1.00
44.67
C


ATOM
12050
CB
LEU
B
258
−60.332
−9.130
−22.438
1.00
44.61
C


ATOM
12053
CG
LEU
B
258
−60.422
−9.323
−20.920
1.00
43.83
C


ATOM
12055
CD1
LEU
B
258
−59.045
−9.552
−20.339
1.00
43.48
C


ATOM
12059
CD2
LEU
B
258
−61.073
−8.139
−20.262
1.00
43.85
C


ATOM
12063
C
LEU
B
258
−61.347
−11.286
−23.090
1.00
43.49
C


ATOM
12064
O
LEU
B
258
−61.375
−12.220
−22.295
1.00
43.84
O


ATOM
12066
N
ILE
B
259
−62.364
−10.968
−23.883
1.00
41.92
N


ATOM
12067
CA
ILE
B
259
−63.617
−11.711
−23.841
1.00
40.80
C


ATOM
12069
CB
ILE
B
259
−64.718
−11.132
−24.783
1.00
41.05
C


ATOM
12071
CG1
ILE
B
259
−64.852
−9.598
−24.653
1.00
41.61
C


ATOM
12074
CD1
ILE
B
259
−65.200
−8.874
−25.985
1.00
42.16
C


ATOM
12078
CG2
ILE
B
259
−66.065
−11.816
−24.496
1.00
40.46
C


ATOM
12082
C
ILE
B
259
−63.319
−13.151
−24.250
1.00
39.50
C


ATOM
12083
O
ILE
B
259
−63.782
−14.082
−23.598
1.00
39.30
O


ATOM
12085
N
GLU
B
260
−62.524
−13.318
−25.316
1.00
37.99
N


ATOM
12086
CA
GLU
B
260
−62.120
−14.652
−25.812
1.00
36.60
C


ATOM
12088
CB
GLU
B
260
−61.329
−14.574
−27.142
1.00
36.75
C


ATOM
12091
CG
GLU
B
260
−62.167
−14.319
−28.425
1.00
37.79
C


ATOM
12094
CD
GLU
B
260
−62.134
−12.851
−28.905
1.00
39.47
C


ATOM
12095
OE1
GLU
B
260
−62.207
−11.923
−28.055
1.00
39.97
O


ATOM
12096
OE2
GLU
B
260
−62.033
−12.628
−30.139
1.00
40.31
O


ATOM
12097
C
GLU
B
260
−61.282
−15.381
−24.770
1.00
34.83
C


ATOM
12098
O
GLU
B
260
−61.465
−16.568
−24.544
1.00
34.69
O


ATOM
12100
N
SER
B
261
−60.365
−14.669
−24.131
1.00
32.89
N


ATOM
12101
CA
SER
B
261
−59.508
−15.292
−23.139
1.00
31.48
C


ATOM
12103
CB
SER
B
261
−58.341
−14.376
−22.768
1.00
31.55
C


ATOM
12106
OG
SER
B
261
−57.298
−14.493
−23.728
1.00
31.75
O


ATOM
12108
C
SER
B
261
−60.294
−15.716
−21.900
1.00
30.04
C


ATOM
12109
O
SER
B
261
−59.921
−16.674
−21.208
1.00
29.70
O


ATOM
12111
N
PHE
B
262
−61.389
−15.021
−21.618
1.00
28.25
N


ATOM
12112
CA
PHE
B
262
−62.203
−15.404
−20.480
1.00
26.75
C


ATOM
12114
CB
PHE
B
262
−63.111
−14.278
−20.009
1.00
26.36
C


ATOM
12117
CG
PHE
B
262
−63.532
−14.453
−18.603
1.00
24.33
C


ATOM
12118
CD1
PHE
B
262
−62.697
−14.092
−17.584
1.00
22.66
C


ATOM
12120
CE1
PHE
B
262
−63.047
−14.283
−16.301
1.00
22.65
C


ATOM
12122
CZ
PHE
B
262
−64.245
−14.873
−16.007
1.00
23.93
C


ATOM
12124
CE2
PHE
B
262
−65.076
−15.267
−17.010
1.00
24.08
C


ATOM
12126
CD2
PHE
B
262
−64.711
−15.067
−18.304
1.00
23.96
C


ATOM
12128
C
PHE
B
262
−63.017
−16.660
−20.751
1.00
26.02
C


ATOM
12129
O
PHE
B
262
−63.074
−17.546
−19.909
1.00
26.32
O


ATOM
12131
N
TYR
B
263
−63.652
−16.732
−21.915
1.00
24.98
N


ATOM
12132
CA
TYR
B
263
−64.299
−17.969
−22.396
1.00
24.14
C


ATOM
12134
CB
TYR
B
263
−64.701
−17.756
−23.853
1.00
24.00
C


ATOM
12137
CG
TYR
B
263
−65.080
−18.961
−24.667
1.00
24.32
C


ATOM
12138
CD1
TYR
B
263
−66.302
−19.604
−24.489
1.00
25.03
C


ATOM
12140
CE1
TYR
B
263
−66.666
−20.694
−25.285
1.00
25.81
C


ATOM
12142
CZ
TYR
B
263
−65.798
−21.132
−26.287
1.00
27.06
C


ATOM
12143
OH
TYR
B
263
−66.106
−22.191
−27.123
1.00
27.14
O


ATOM
12145
CE2
TYR
B
263
−64.590
−20.489
−26.479
1.00
26.80
C


ATOM
12147
CD2
TYR
B
263
−64.250
−19.404
−25.680
1.00
25.65
C


ATOM
12149
C
TYR
B
263
−63.351
−19.162
−22.256
1.00
23.35
C


ATOM
12150
O
TYR
B
263
−63.712
−20.219
−21.740
1.00
22.80
O


ATOM
12152
N
TRP
B
264
−62.119
−18.955
−22.702
1.00
22.64
N


ATOM
12153
CA
TRP
B
264
−61.047
−19.918
−22.523
1.00
21.95
C


ATOM
12155
CB
TRP
B
264
−59.737
−19.334
−23.061
1.00
21.84
C


ATOM
12158
CG
TRP
B
264
−58.603
−20.278
−22.964
1.00
22.90
C


ATOM
12159
CD1
TRP
B
264
−57.700
−20.387
−21.943
1.00
23.98
C


ATOM
12161
NE1
TRP
B
264
−56.800
−21.386
−22.214
1.00
23.95
N


ATOM
12163
CE2
TRP
B
264
−57.124
−21.949
−23.417
1.00
23.51
C


ATOM
12164
CD2
TRP
B
264
−58.252
−21.273
−23.915
1.00
23.40
C


ATOM
12165
CE3
TRP
B
264
−58.776
−21.652
−25.147
1.00
23.68
C


ATOM
12167
CZ3
TRP
B
264
−58.178
−22.669
−25.826
1.00
24.44
C


ATOM
12169
CH2
TRP
B
264
−57.060
−23.324
−25.310
1.00
24.97
C


ATOM
12171
CZ2
TRP
B
264
−56.521
−22.978
−24.103
1.00
24.44
C


ATOM
12173
C
TRP
B
264
−60.897
−20.326
−21.050
1.00
21.11
C


ATOM
12174
O
TRP
B
264
−60.768
−21.511
−20.748
1.00
20.98
O


ATOM
12176
N
ALA
B
265
−60.916
−19.344
−20.147
1.00
20.17
N


ATOM
12177
CA
ALA
B
265
−60.774
−19.602
−18.712
1.00
19.36
C


ATOM
12179
CB
ALA
B
265
−60.621
−18.305
−17.944
1.00
19.30
C


ATOM
12183
C
ALA
B
265
−61.947
−20.397
−18.171
1.00
18.63
C


ATOM
12184
O
ALA
B
265
−61.763
−21.349
−17.434
1.00
18.64
O


ATOM
12186
N
VAL
B
266
−63.159
−20.034
−18.557
1.00
18.11
N


ATOM
12187
CA
VAL
B
266
−64.335
−20.758
−18.075
1.00
17.60
C


ATOM
12189
CB
VAL
B
266
−65.652
−20.221
−18.675
1.00
17.38
C


ATOM
12191
CG1
VAL
B
266
−65.902
−18.803
−18.199
1.00
17.09
C


ATOM
12195
CG2
VAL
B
266
−66.817
−21.120
−18.312
1.00
15.40
C


ATOM
12199
C
VAL
B
266
−64.221
−22.246
−18.376
1.00
17.67
C


ATOM
12200
O
VAL
B
266
−64.766
−23.058
−17.647
1.00
18.27
O


ATOM
12202
N
GLY
B
267
−63.516
−22.607
−19.444
1.00
17.43
N


ATOM
12203
CA
GLY
B
267
−63.316
−24.013
−19.789
1.00
16.98
C


ATOM
12206
C
GLY
B
267
−62.253
−24.683
−18.947
1.00
16.55
C


ATOM
12207
O
GLY
B
267
−62.360
−25.859
−18.609
1.00
16.37
O


ATOM
12209
N
VAL
B
268
−61.215
−23.939
−18.608
1.00
16.30
N


ATOM
12210
CA
VAL
B
268
−60.160
−24.500
−17.794
1.00
16.23
C


ATOM
12212
CB
VAL
B
268
−58.893
−23.651
−17.840
1.00
15.68
C


ATOM
12214
CG1
VAL
B
268
−57.857
−24.194
−16.894
1.00
15.01
C


ATOM
12218
CG2
VAL
B
268
−58.361
−23.674
−19.235
1.00
15.01
C


ATOM
12222
C
VAL
B
268
−60.654
−24.718
−16.374
1.00
16.86
C


ATOM
12223
O
VAL
B
268
−60.421
−25.784
−15.813
1.00
16.93
O


ATOM
12225
N
ALA
B
269
−61.362
−23.733
−15.817
1.00
17.64
N


ATOM
12226
CA
ALA
B
269
−61.911
−23.838
−14.461
1.00
18.46
C


ATOM
12228
CB
ALA
B
269
−60.958
−23.215
−13.472
1.00
18.15
C


ATOM
12232
C
ALA
B
269
−63.312
−23.213
−14.340
1.00
19.34
C


ATOM
12233
O
ALA
B
269
−63.448
−22.030
−14.047
1.00
19.78
O


ATOM
12235
N
PHE
B
270
−64.347
−24.028
−14.529
1.00
20.34
N


ATOM
12236
CA
PHE
B
270
−65.724
−23.535
−14.631
1.00
21.21
C


ATOM
12238
CB
PHE
B
270
−66.564
−24.483
−15.482
1.00
21.34
C


ATOM
12241
CG
PHE
B
270
−67.083
−25.667
−14.718
1.00
22.46
C


ATOM
12242
CD1
PHE
B
270
−68.294
−25.596
−14.039
1.00
23.19
C


ATOM
12244
CE1
PHE
B
270
−68.756
−26.672
−13.315
1.00
23.27
C


ATOM
12246
CZ
PHE
B
270
−68.003
−27.836
−13.258
1.00
23.35
C


ATOM
12248
CE2
PHE
B
270
−66.794
−27.918
−13.932
1.00
22.73
C


ATOM
12250
CD2
PHE
B
270
−66.340
−26.843
−14.646
1.00
22.88
C


ATOM
12252
C
PHE
B
270
−66.464
−23.366
−13.316
1.00
21.98
C


ATOM
12253
O
PHE
B
270
−67.408
−22.583
−13.268
1.00
21.93
O


ATOM
12255
N
GLU
B
271
−66.102
−24.126
−12.275
1.00
23.01
N


ATOM
12256
CA
GLU
B
271
−66.929
−24.142
−11.037
1.00
23.99
C


ATOM
12258
CB
GLU
B
271
−66.679
−25.353
−10.113
1.00
24.25
C


ATOM
12261
CG
GLU
B
271
−65.286
−25.877
−10.092
1.00
26.22
C


ATOM
12264
CD
GLU
B
271
−65.013
−26.914
−11.183
1.00
29.01
C


ATOM
12265
OE1
GLU
B
271
−65.563
−28.032
−11.090
1.00
31.36
O


ATOM
12266
OE2
GLU
B
271
−64.232
−26.622
−12.121
1.00
30.98
O


ATOM
12267
C
GLU
B
271
−66.795
−22.831
−10.281
1.00
23.90
C


ATOM
12268
O
GLU
B
271
−65.702
−22.296
−10.181
1.00
24.29
O


ATOM
12270
N
PRO
B
272
−67.913
−22.316
−9.745
1.00
24.09
N


ATOM
12271
CA
PRO
B
272
−68.005
−20.885
−9.428
1.00
24.11
C


ATOM
12273
CB
PRO
B
272
−69.380
−20.751
−8.751
1.00
24.03
C


ATOM
12276
CG
PRO
B
272
−70.083
−22.069
−8.997
1.00
24.12
C


ATOM
12279
CD
PRO
B
272
−69.013
−23.083
−9.132
1.00
23.99
C


ATOM
12282
C
PRO
B
272
−66.900
−20.386
−8.501
1.00
24.28
C


ATOM
12283
O
PRO
B
272
−66.421
−19.261
−8.676
1.00
24.40
O


ATOM
12284
N
GLN
B
273
−66.480
−21.222
−7.547
1.00
24.20
N


ATOM
12285
CA
GLN
B
273
−65.500
−20.809
−6.551
1.00
24.20
C


ATOM
12287
CB
GLN
B
273
−65.437
−21.840
−5.426
1.00
24.42
C


ATOM
12290
CG
GLN
B
273
−64.750
−23.176
−5.775
1.00
25.17
C


ATOM
12293
CD
GLN
B
273
−65.717
−24.334
−6.007
1.00
26.13
C


ATOM
12294
OE1
GLN
B
273
−66.857
−24.141
−6.453
1.00
27.46
O


ATOM
12295
NE2
GLN
B
273
−65.251
−25.554
−5.721
1.00
25.05
N


ATOM
12298
C
GLN
B
273
−64.086
−20.546
−7.088
1.00
24.27
C


ATOM
12299
O
GLN
B
273
−63.221
−20.151
−6.329
1.00
24.32
O


ATOM
12301
N
TYR
B
274
−63.842
−20.759
−8.380
1.00
24.59
N


ATOM
12302
CA
TYR
B
274
−62.509
−20.551
−8.962
1.00
24.85
C


ATOM
12304
CB
TYR
B
274
−62.129
−21.724
−9.880
1.00
24.62
C


ATOM
12307
CG
TYR
B
274
−62.009
−23.054
−9.184
1.00
24.35
C


ATOM
12308
CD1
TYR
B
274
−61.395
−23.169
−7.946
1.00
24.62
C


ATOM
12310
CE1
TYR
B
274
−61.280
−24.391
−7.314
1.00
25.10
C


ATOM
12312
CZ
TYR
B
274
−61.771
−25.525
−7.920
1.00
24.92
C


ATOM
12313
OH
TYR
B
274
−61.661
−26.748
−7.299
1.00
24.73
O


ATOM
12315
CE2
TYR
B
274
−62.366
−25.437
−9.150
1.00
24.76
C


ATOM
12317
CD2
TYR
B
274
−62.477
−24.203
−9.779
1.00
24.70
C


ATOM
12319
C
TYR
B
274
−62.409
−19.235
−9.745
1.00
25.27
C


ATOM
12320
O
TYR
B
274
−61.956
−19.210
−10.895
1.00
25.42
O


ATOM
12322
N
SER
B
275
−62.808
−18.134
−9.121
1.00
25.40
N


ATOM
12323
CA
SER
B
275
−62.755
−16.849
−9.802
1.00
25.43
C


ATOM
12325
CB
SER
B
275
−63.615
−15.814
−9.071
1.00
25.53
C


ATOM
12328
OG
SER
B
275
−65.006
−16.068
−9.291
1.00
25.52
O


ATOM
12330
C
SER
B
275
−61.300
−16.403
−9.948
1.00
25.36
C


ATOM
12331
O
SER
B
275
−60.885
−15.903
−10.996
1.00
25.17
O


ATOM
12333
N
ASP
B
276
−60.517
−16.617
−8.901
1.00
25.41
N


ATOM
12334
CA
ASP
B
276
−59.095
−16.353
−8.979
1.00
25.42
C


ATOM
12336
CB
ASP
B
276
−58.390
−16.794
−7.712
1.00
25.44
C


ATOM
12339
CG
ASP
B
276
−58.728
−15.925
−6.558
1.00
26.32
C


ATOM
12340
OD1
ASP
B
276
−59.302
−14.857
−6.818
1.00
27.50
O


ATOM
12341
OD2
ASP
B
276
−58.442
−16.299
−5.398
1.00
30.16
O


ATOM
12342
C
ASP
B
276
−58.495
−17.069
−10.157
1.00
25.20
C


ATOM
12343
O
ASP
B
276
−57.745
−16.461
−10.929
1.00
25.66
O


ATOM
12345
N
CYS
B
277
−58.814
−18.353
−10.305
1.00
24.60
N


ATOM
12346
CA
CYS
B
277
−58.228
−19.110
−11.391
1.00
24.30
C


ATOM
12348
CB
CYS
B
277
−58.684
−20.561
−11.392
1.00
24.28
C


ATOM
12351
SG
CYS
B
277
−57.737
−21.574
−12.569
1.00
23.82
S


ATOM
12353
C
CYS
B
277
−58.593
−18.448
−12.698
1.00
24.09
C


ATOM
12354
O
CYS
B
277
−57.727
−18.013
−13.450
1.00
23.87
O


ATOM
12356
N
ARG
B
278
−59.893
−18.329
−12.933
1.00
24.22
N


ATOM
12357
CA
ARG
B
278
−60.400
−17.746
−14.171
1.00
23.98
C


ATOM
12359
CB
ARG
B
278
−61.917
−17.556
−14.123
1.00
23.93
C


ATOM
12362
CG
ARG
B
278
−62.662
−18.871
−14.283
1.00
24.07
C


ATOM
12365
CD
ARG
B
278
−64.132
−18.670
−14.607
1.00
24.48
C


ATOM
12368
NE
ARG
B
278
−64.874
−18.141
−13.472
1.00
23.75
N


ATOM
12370
CZ
ARG
B
278
−65.254
−18.856
−12.419
1.00
23.72
C


ATOM
12371
NH1
ARG
B
278
−64.973
−20.149
−12.326
1.00
22.86
N


ATOM
12374
NH2
ARG
B
278
−65.927
−18.262
−11.443
1.00
25.08
N


ATOM
12377
C
ARG
B
278
−59.701
−16.453
−14.491
1.00
23.56
C


ATOM
12378
O
ARG
B
278
−59.296
−16.265
−15.628
1.00
23.87
O


ATOM
12380
N
ASN
B
279
−59.512
−15.593
−13.495
1.00
23.24
N


ATOM
12381
CA
ASN
B
279
−58.854
−14.301
−13.734
1.00
23.48
C


ATOM
12383
CB
ASN
B
279
−59.055
−13.340
−12.574
1.00
24.05
C


ATOM
12386
CG
ASN
B
279
−60.510
−13.017
−12.346
1.00
26.58
C


ATOM
12387
OD1
ASN
B
279
−61.378
−13.310
−13.189
1.00
28.96
O


ATOM
12388
ND2
ASN
B
279
−60.799
−12.416
−11.194
1.00
30.30
N


ATOM
12391
C
ASN
B
279
−57.376
−14.404
−14.027
1.00
22.58
C


ATOM
12392
O
ASN
B
279
−56.903
−13.749
−14.941
1.00
22.73
O


ATOM
12394
N
SER
B
280
−56.648
−15.205
−13.251
1.00
21.62
N


ATOM
12395
CA
SER
B
280
−55.243
−15.483
−13.550
1.00
20.73
C


ATOM
12397
CB
SER
B
280
−54.696
−16.597
−12.659
1.00
20.85
C


ATOM
12400
OG
SER
B
280
−53.805
−16.099
−11.683
1.00
21.55
O


ATOM
12402
C
SER
B
280
−55.091
−15.896
−15.004
1.00
20.04
C


ATOM
12403
O
SER
B
280
−54.293
−15.312
−15.738
1.00
19.87
O


ATOM
12405
N
VAL
B
281
−55.875
−16.893
−15.413
1.00
19.24
N


ATOM
12406
CA
VAL
B
281
−55.720
−17.491
−16.720
1.00
18.77
C


ATOM
12408
CB
VAL
B
281
−56.482
−18.834
−16.815
1.00
18.74
C


ATOM
12410
CG1
VAL
B
281
−56.467
−19.404
−18.251
1.00
18.34
C


ATOM
12414
CG2
VAL
B
281
−55.863
−19.844
−15.848
1.00
18.13
C


ATOM
12418
C
VAL
B
281
−56.142
−16.486
−17.787
1.00
18.89
C


ATOM
12419
O
VAL
B
281
−55.483
−16.364
−18.832
1.00
18.75
O


ATOM
12421
N
ALA
B
282
−57.210
−15.739
−17.505
1.00
18.85
N


ATOM
12422
CA
ALA
B
282
−57.709
−14.723
−18.439
1.00
18.80
C


ATOM
12424
CB
ALA
B
282
−59.051
−14.191
−17.991
1.00
18.32
C


ATOM
12428
C
ALA
B
282
−56.718
−13.576
−18.615
1.00
18.94
C


ATOM
12429
O
ALA
B
282
−56.582
−13.021
−19.700
1.00
19.16
O


ATOM
12431
N
LYS
B
283
−56.022
−13.211
−17.553
1.00
19.24
N


ATOM
12432
CA
LYS
B
283
−55.037
−12.145
−17.658
1.00
19.62
C


ATOM
12434
CB
LYS
B
283
−54.649
−11.615
−16.276
1.00
19.68
C


ATOM
12437
CG
LYS
B
283
−55.779
−10.923
−15.518
1.00
19.95
C


ATOM
12440
CD
LYS
B
283
−55.265
−10.398
−14.196
1.00
21.72
C


ATOM
12443
CE
LYS
B
283
−56.364
−10.236
−13.164
1.00
23.83
C


ATOM
12446
NZ
LYS
B
283
−56.129
−9.003
−12.351
1.00
24.72
N


ATOM
12450
C
LYS
B
283
−53.802
−12.643
−18.396
1.00
19.77
C


ATOM
12451
O
LYS
B
283
−53.237
−11.929
−19.208
1.00
20.05
O


ATOM
12453
N
MET
B
284
−53.387
−13.871
−18.117
1.00
19.88
N


ATOM
12454
CA
MET
B
284
−52.136
−14.367
−18.652
1.00
19.90
C


ATOM
12456
CB
MET
B
284
−51.719
−15.676
−17.960
1.00
20.08
C


ATOM
12459
CG
MET
B
284
−51.125
−15.505
−16.560
1.00
20.62
C


ATOM
12462
SD
MET
B
284
−49.782
−14.295
−16.485
1.00
23.69
S


ATOM
12463
CE
MET
B
284
−50.600
−12.872
−15.754
1.00
23.82
C


ATOM
12467
C
MET
B
284
−52.257
−14.572
−20.145
1.00
19.82
C


ATOM
12468
O
MET
B
284
−51.326
−14.269
−20.889
1.00
20.21
O


ATOM
12470
N
PHE
B
285
−53.401
−15.088
−20.579
1.00
19.72
N


ATOM
12471
CA
PHE
B
285
−53.620
−15.428
−21.986
1.00
19.66
C


ATOM
12473
CB
PHE
B
285
−54.863
−16.324
−22.114
1.00
20.09
C


ATOM
12476
CG
PHE
B
285
−54.986
−17.077
−23.427
1.00
21.18
C


ATOM
12477
CD1
PHE
B
285
−53.877
−17.362
−24.232
1.00
22.86
C


ATOM
12479
CE1
PHE
B
285
−54.013
−18.070
−25.424
1.00
22.58
C


ATOM
12481
CZ
PHE
B
285
−55.254
−18.518
−25.806
1.00
23.49
C


ATOM
12483
CE2
PHE
B
285
−56.365
−18.254
−25.002
1.00
23.36
C


ATOM
12485
CD2
PHE
B
285
−56.222
−17.551
−23.823
1.00
21.76
C


ATOM
12487
C
PHE
B
285
−53.781
−14.148
−22.781
1.00
19.18
C


ATOM
12488
O
PHE
B
285
−53.333
−14.046
−23.914
1.00
18.98
O


ATOM
12490
N
SER
B
286
−54.410
−13.161
−22.164
1.00
18.92
N


ATOM
12491
CA
SER
B
286
−54.488
−11.843
−22.753
1.00
19.15
C


ATOM
12493
CB
SER
B
286
−55.292
−10.894
−21.863
1.00
19.38
C


ATOM
12496
OG
SER
B
286
−56.684
−11.180
−21.957
1.00
20.24
O


ATOM
12498
C
SER
B
286
−53.106
−11.267
−23.032
1.00
18.91
C


ATOM
12499
O
SER
B
286
−52.894
−10.668
−24.087
1.00
19.48
O


ATOM
12501
N
PHE
B
287
−52.172
−11.446
−22.102
1.00
18.48
N


ATOM
12502
CA
PHE
B
287
−50.768
−11.065
−22.339
1.00
18.30
C


ATOM
12504
CB
PHE
B
287
−49.958
−11.035
−21.044
1.00
18.27
C


ATOM
12507
CG
PHE
B
287
−50.040
−9.740
−20.324
1.00
18.59
C


ATOM
12508
CD1
PHE
B
287
−49.375
−8.624
−20.813
1.00
20.02
C


ATOM
12510
CE1
PHE
B
287
−49.456
−7.410
−20.152
1.00
20.57
C


ATOM
12512
CZ
PHE
B
287
−50.208
−7.308
−18.985
1.00
20.02
C


ATOM
12514
CE2
PHE
B
287
−50.869
−8.417
−18.503
1.00
19.78
C


ATOM
12516
CD2
PHE
B
287
−50.781
−9.623
−19.164
1.00
19.05
C


ATOM
12518
C
PHE
B
287
−50.067
−11.978
−23.339
1.00
18.01
C


ATOM
12519
O
PHE
B
287
−49.374
−11.498
−24.229
1.00
18.00
O


ATOM
12521
N
VAL
B
288
−50.236
−13.286
−23.194
1.00
17.69
N


ATOM
12522
CA
VAL
B
288
−49.720
−14.206
−24.197
1.00
17.66
C


ATOM
12524
CB
VAL
B
288
−50.168
−15.649
−23.930
1.00
17.37
C


ATOM
12526
CG1
VAL
B
288
−49.937
−16.529
−25.134
1.00
16.24
C


ATOM
12530
CG2
VAL
B
288
−49.414
−16.185
−22.758
1.00
17.21
C


ATOM
12534
C
VAL
B
288
−50.081
−13.761
−25.633
1.00
18.08
C


ATOM
12535
O
VAL
B
288
−49.207
−13.710
−26.492
1.00
18.08
O


ATOM
12537
N
THR
B
289
−51.327
−13.396
−25.895
1.00
18.42
N


ATOM
12538
CA
THR
B
289
−51.683
−13.001
−27.256
1.00
19.33
C


ATOM
12540
CB
THR
B
289
−53.171
−12.736
−27.401
1.00
19.35
C


ATOM
12542
OG1
THR
B
289
−53.573
−11.895
−26.321
1.00
21.59
O


ATOM
12544
CG2
THR
B
289
−53.973
−14.046
−27.350
1.00
19.09
C


ATOM
12548
C
THR
B
289
−50.901
−11.766
−27.723
1.00
19.71
C


ATOM
12549
O
THR
B
289
−50.469
−11.707
−28.891
1.00
19.88
O


ATOM
12551
N
ILE
B
290
−50.689
−10.796
−26.829
1.00
19.94
N


ATOM
12552
CA
ILE
B
290
−49.943
−9.585
−27.225
1.00
20.39
C


ATOM
12554
CB
ILE
B
290
−50.069
−8.410
−26.221
1.00
20.50
C


ATOM
12556
CG1
ILE
B
290
−51.510
−8.137
−25.823
1.00
21.25
C


ATOM
12559
CD1
ILE
B
290
−51.650
−6.854
−25.002
1.00
21.58
C


ATOM
12563
CG2
ILE
B
290
−49.546
−7.134
−26.853
1.00
20.39
C


ATOM
12567
C
ILE
B
290
−48.434
−9.840
−27.438
1.00
20.45
C


ATOM
12568
O
ILE
B
290
−47.853
−9.317
−28.393
1.00
20.76
O


ATOM
12570
N
ILE
B
291
−47.808
−10.616
−26.546
1.00
20.21
N


ATOM
12571
CA
ILE
B
291
−46.375
−10.869
−26.618
1.00
19.87
C


ATOM
12573
CB
ILE
B
291
−45.840
−11.603
−25.407
1.00
19.79
C


ATOM
12575
CG1
ILE
B
291
−46.205
−10.879
−24.102
1.00
19.71
C


ATOM
12578
CD1
ILE
B
291
−45.326
−9.734
−23.748
1.00
19.77
C


ATOM
12582
CG2
ILE
B
291
−44.336
−11.747
−25.516
1.00
18.91
C


ATOM
12586
C
ILE
B
291
−46.134
−11.735
−27.825
1.00
20.44
C


ATOM
12587
O
ILE
B
291
−45.233
−11.481
−28.600
1.00
20.35
O


ATOM
12589
N
ASP
B
292
−46.965
−12.751
−28.005
1.00
21.33
N


ATOM
12590
CA
ASP
B
292
−46.915
−13.563
−29.228
1.00
22.13
C


ATOM
12592
CB
ASP
B
292
−48.120
−14.508
−29.316
1.00
22.40
C


ATOM
12595
CG
ASP
B
292
−47.847
−15.741
−30.160
1.00
23.47
C


ATOM
12596
OD1
ASP
B
292
−46.852
−15.765
−30.923
1.00
24.83
O


ATOM
12597
OD2
ASP
B
292
−48.637
−16.707
−30.038
1.00
25.22
O


ATOM
12598
C
ASP
B
292
−46.856
−12.716
−30.507
1.00
22.28
C


ATOM
12599
O
ASP
B
292
−46.088
−13.033
−31.399
1.00
22.73
O


ATOM
12601
N
ASP
B
293
−47.656
−11.658
−30.611
1.00
22.22
N


ATOM
12602
CA
ASP
B
293
−47.630
−10.828
−31.822
1.00
22.43
C


ATOM
12604
CB
ASP
B
293
−48.840
−9.894
−31.885
1.00
23.03
C


ATOM
12607
CG
ASP
B
293
−50.157
−10.644
−31.959
1.00
24.69
C


ATOM
12608
OD1
ASP
B
293
−50.113
−11.900
−32.025
1.00
28.66
O


ATOM
12609
OD2
ASP
B
293
−51.231
−9.986
−31.930
1.00
23.57
O


ATOM
12610
C
ASP
B
293
−46.354
−9.997
−31.921
1.00
21.94
C


ATOM
12611
O
ASP
B
293
−45.898
−9.674
−33.019
1.00
22.31
O


ATOM
12613
N
ILE
B
294
−45.783
−9.643
−30.780
1.00
21.05
N


ATOM
12614
CA
ILE
B
294
−44.513
−8.941
−30.770
1.00
20.53
C


ATOM
12616
CB
ILE
B
294
−44.166
−8.448
−29.344
1.00
20.42
C


ATOM
12618
CG1
ILE
B
294
−45.150
−7.349
−28.945
1.00
19.66
C


ATOM
12621
CD1
ILE
B
294
−45.057
−6.956
−27.536
1.00
18.81
C


ATOM
12625
CG2
ILE
B
294
−42.726
−7.924
−29.253
1.00
19.99
C


ATOM
12629
C
ILE
B
294
−43.416
−9.845
−31.343
1.00
20.44
C


ATOM
12630
O
ILE
B
294
−42.650
−9.431
−32.205
1.00
20.17
O


ATOM
12632
N
TYR
B
295
−43.355
−11.088
−30.886
1.00
20.65
N


ATOM
12633
CA
TYR
B
295
−42.313
−12.008
−31.353
1.00
20.76
C


ATOM
12635
CB
TYR
B
295
−42.145
−13.198
−30.412
1.00
20.29
C


ATOM
12638
CG
TYR
B
295
−41.393
−12.926
−29.129
1.00
17.92
C


ATOM
12639
CD1
TYR
B
295
−40.117
−13.409
−28.936
1.00
17.62
C


ATOM
12641
CE1
TYR
B
295
−39.427
−13.195
−27.747
1.00
17.12
C


ATOM
12643
CZ
TYR
B
295
−40.022
−12.493
−26.730
1.00
16.61
C


ATOM
12644
OH
TYR
B
295
−39.357
−12.281
−25.528
1.00
14.46
O


ATOM
12646
CE2
TYR
B
295
−41.296
−12.008
−26.911
1.00
16.82
C


ATOM
12648
CD2
TYR
B
295
−41.972
−12.235
−28.102
1.00
16.39
C


ATOM
12650
C
TYR
B
295
−42.619
−12.519
−32.749
1.00
21.65
C


ATOM
12651
O
TYR
B
295
−41.698
−12.751
−33.540
1.00
22.13
O


ATOM
12653
N
ASP
B
296
−43.905
−12.677
−33.049
1.00
22.47
N


ATOM
12654
CA
ASP
B
296
−44.327
−13.316
−34.285
1.00
23.52
C


ATOM
12656
CB
ASP
B
296
−45.761
−13.851
−34.185
1.00
23.99
C


ATOM
12659
CG
ASP
B
296
−46.134
−14.744
−35.369
1.00
26.22
C


ATOM
12660
OD1
ASP
B
296
−45.508
−15.832
−35.538
1.00
27.70
O


ATOM
12661
OD2
ASP
B
296
−47.052
−14.348
−36.128
1.00
28.78
O


ATOM
12662
C
ASP
B
296
−44.228
−12.392
−35.478
1.00
23.68
C


ATOM
12663
O
ASP
B
296
−43.673
−12.784
−36.502
1.00
24.01
O


ATOM
12665
N
VAL
B
297
−44.765
−11.178
−35.364
1.00
23.90
N


ATOM
12666
CA
VAL
B
297
−44.801
−10.273
−36.524
1.00
23.99
C


ATOM
12668
CB
VAL
B
297
−46.251
−10.003
−36.973
1.00
23.87
C


ATOM
12670
CG1
VAL
B
297
−46.949
−11.310
−37.197
1.00
23.75
C


ATOM
12674
CG2
VAL
B
297
−47.002
−9.136
−35.966
1.00
23.05
C


ATOM
12678
C
VAL
B
297
−44.060
−8.940
−36.383
1.00
24.27
C


ATOM
12679
O
VAL
B
297
−43.485
−8.465
−37.342
1.00
23.84
O


ATOM
12681
N
TYR
B
298
−44.058
−8.347
−35.199
1.00
24.87
N


ATOM
12682
CA
TYR
B
298
−43.760
−6.926
−35.078
1.00
25.67
C


ATOM
12684
CB
TYR
B
298
−44.656
−6.294
−34.010
1.00
26.06
C


ATOM
12687
CG
TYR
B
298
−44.520
−4.789
−33.932
1.00
27.95
C


ATOM
12688
CD1
TYR
B
298
−45.097
−3.968
−34.901
1.00
29.74
C


ATOM
12690
CE1
TYR
B
298
−44.982
−2.574
−34.843
1.00
29.93
C


ATOM
12692
CZ
TYR
B
298
−44.282
−1.991
−33.806
1.00
29.96
C


ATOM
12693
OH
TYR
B
298
−44.165
−.631
−33.746
1.00
30.01
O


ATOM
12695
CE2
TYR
B
298
−43.697
−2.777
−32.829
1.00
30.39
C


ATOM
12697
CD2
TYR
B
298
−43.811
−4.181
−32.899
1.00
29.96
C


ATOM
12699
C
TYR
B
298
−42.318
−6.592
−34.755
1.00
25.76
C


ATOM
12700
O
TYR
B
298
−41.696
−5.830
−35.470
1.00
26.54
O


ATOM
12702
N
GLY
B
299
−41.798
−7.111
−33.653
1.00
25.89
N


ATOM
12703
CA
GLY
B
299
−40.473
−6.706
−33.167
1.00
25.78
C


ATOM
12706
C
GLY
B
299
−39.346
−7.341
−33.951
1.00
25.52
C


ATOM
12707
O
GLY
B
299
−39.513
−8.410
−34.525
1.00
26.04
O


ATOM
12709
N
THR
B
300
−38.196
−6.685
−33.990
1.00
25.32
N


ATOM
12710
CA
THR
B
300
−37.063
−7.237
−34.716
1.00
25.16
C


ATOM
12712
CB
THR
B
300
−36.088
−6.164
−35.295
1.00
25.08
C


ATOM
12714
OG1
THR
B
300
−35.191
−5.710
−34.281
1.00
25.07
O


ATOM
12716
CG2
THR
B
300
−36.833
−4.978
−35.875
1.00
25.08
C


ATOM
12720
C
THR
B
300
−36.324
−8.139
−33.771
1.00
25.13
C


ATOM
12721
O
THR
B
300
−36.335
−7.950
−32.570
1.00
25.28
O


ATOM
12723
N
LEU
B
301
−35.662
−9.129
−34.332
1.00
25.50
N


ATOM
12724
CA
LEU
B
301
−34.918
−10.093
−33.538
1.00
25.42
C


ATOM
12726
CB
LEU
B
301
−34.148
−11.020
−34.479
1.00
25.17
C


ATOM
12729
CG
LEU
B
301
−33.624
−12.338
−33.941
1.00
24.95
C


ATOM
12731
CD1
LEU
B
301
−34.715
−13.233
−33.437
1.00
24.42
C


ATOM
12735
CD2
LEU
B
301
−32.900
−12.998
−35.080
1.00
26.50
C


ATOM
12739
C
LEU
B
301
−33.995
−9.425
−32.485
1.00
25.54
C


ATOM
12740
O
LEU
B
301
−33.843
−9.959
−31.394
1.00
25.35
O


ATOM
12742
N
ASP
B
302
−33.417
−8.257
−32.782
1.00
25.68
N


ATOM
12743
CA
ASP
B
302
−32.598
−7.550
−31.774
1.00
26.02
C


ATOM
12745
CB
ASP
B
302
−31.800
−6.377
−32.378
1.00
26.37
C


ATOM
12748
CG
ASP
B
302
−30.563
−6.833
−33.167
1.00
28.27
C


ATOM
12749
OD1
ASP
B
302
−29.508
−6.156
−33.048
1.00
29.05
O


ATOM
12750
OD2
ASP
B
302
−30.652
−7.852
−33.910
1.00
31.04
O


ATOM
12751
C
ASP
B
302
−33.472
−7.037
−30.630
1.00
25.69
C


ATOM
12752
O
ASP
B
302
−33.110
−7.142
−29.447
1.00
25.90
O


ATOM
12754
N
GLU
B
303
−34.622
−6.473
−30.993
1.00
25.04
N


ATOM
12755
CA
GLU
B
303
−35.600
−6.029
−30.011
1.00
24.33
C


ATOM
12757
CB
GLU
B
303
−36.783
−5.346
−30.697
1.00
24.20
C


ATOM
12760
CG
GLU
B
303
−36.399
−4.109
−31.466
1.00
24.23
C


ATOM
12763
CD
GLU
B
303
−37.589
−3.382
−32.063
1.00
24.23
C


ATOM
12764
OE1
GLU
B
303
−38.496
−4.065
−32.604
1.00
22.66
O


ATOM
12765
OE2
GLU
B
303
−37.592
−2.121
−31.995
1.00
23.93
O


ATOM
12766
C
GLU
B
303
−36.103
−7.212
−29.193
1.00
23.87
C


ATOM
12767
O
GLU
B
303
−36.396
−7.058
−28.011
1.00
23.59
O


ATOM
12769
N
LEU
B
304
−36.204
−8.383
−29.825
1.00
23.39
N


ATOM
12770
CA
LEU
B
304
−36.766
−9.570
−29.166
1.00
23.23
C


ATOM
12772
CB
LEU
B
304
−37.180
−10.624
−30.208
1.00
22.83
C


ATOM
12775
CG
LEU
B
304
−38.381
−10.217
−31.074
1.00
22.06
C


ATOM
12777
CD1
LEU
B
304
−38.822
−11.321
−32.037
1.00
20.11
C


ATOM
12781
CD2
LEU
B
304
−39.551
−9.800
−30.163
1.00
21.94
C


ATOM
12785
C
LEU
B
304
−35.810
−10.147
−28.106
1.00
23.37
C


ATOM
12786
O
LEU
B
304
−36.248
−10.785
−27.141
1.00
23.22
O


ATOM
12788
N
GLU
B
305
−34.514
−9.892
−28.283
1.00
23.30
N


ATOM
12789
CA
GLU
B
305
−33.512
−10.308
−27.324
1.00
23.18
C


ATOM
12791
CB
GLU
B
305
−32.114
−10.176
−27.899
1.00
23.58
C


ATOM
12794
CG
GLU
B
305
−31.832
−11.142
−29.047
1.00
25.27
C


ATOM
12797
CD
GLU
B
305
−31.306
−12.488
−28.585
1.00
27.91
C


ATOM
12798
OE1
GLU
B
305
−30.946
−13.303
−29.470
1.00
29.87
O


ATOM
12799
OE2
GLU
B
305
−31.244
−12.728
−27.351
1.00
29.28
O


ATOM
12800
C
GLU
B
305
−33.633
−9.440
−26.104
1.00
22.52
C


ATOM
12801
O
GLU
B
305
−33.793
−9.956
−25.005
1.00
23.39
O


ATOM
12803
N
LEU
B
306
−33.576
−8.124
−26.274
1.00
21.51
N


ATOM
12804
CA
LEU
B
306
−33.715
−7.224
−25.114
1.00
20.70
C


ATOM
12806
CB
LEU
B
306
−33.899
−5.764
−25.544
1.00
20.59
C


ATOM
12809
CG
LEU
B
306
−32.769
−5.065
−26.293
1.00
20.12
C


ATOM
12811
CD1
LEU
B
306
−32.992
−3.576
−26.203
1.00
19.89
C


ATOM
12815
CD2
LEU
B
306
−31.402
−5.437
−25.739
1.00
19.81
C


ATOM
12819
C
LEU
B
306
−34.896
−7.645
−24.237
1.00
19.91
C


ATOM
12820
O
LEU
B
306
−34.771
−7.778
−23.031
1.00
19.53
O


ATOM
12822
N
PHE
B
307
−36.030
−7.883
−24.872
1.00
19.48
N


ATOM
12823
CA
PHE
B
307
−37.231
−8.267
−24.170
1.00
19.36
C


ATOM
12825
CB
PHE
B
307
−38.400
−8.406
−25.138
1.00
19.54
C


ATOM
12828
CG
PHE
B
307
−39.729
−8.296
−24.482
1.00
19.66
C


ATOM
12829
CD1
PHE
B
307
−40.365
−7.079
−24.400
1.00
21.03
C


ATOM
12831
CE1
PHE
B
307
−41.590
−6.968
−23.787
1.00
21.62
C


ATOM
12833
CZ
PHE
B
307
−42.182
−8.072
−23.248
1.00
20.25
C


ATOM
12835
CE2
PHE
B
307
−41.553
−9.292
−23.329
1.00
20.07
C


ATOM
12837
CD2
PHE
B
307
−40.338
−9.400
−23.939
1.00
19.62
C


ATOM
12839
C
PHE
B
307
−37.039
−9.577
−23.443
1.00
19.22
C


ATOM
12840
O
PHE
B
307
−37.354
−9.684
−22.260
1.00
19.50
O


ATOM
12842
N
THR
B
308
−36.532
−10.583
−24.143
1.00
18.96
N


ATOM
12843
CA
THR
B
308
−36.321
−11.874
−23.511
1.00
18.84
C


ATOM
12845
CB
THR
B
308
−35.774
−12.879
−24.490
1.00
18.62
C


ATOM
12847
OG1
THR
B
308
−36.687
−12.988
−25.579
1.00
18.88
O


ATOM
12849
CG2
THR
B
308
−35.626
−14.228
−23.833
1.00
18.74
C


ATOM
12853
C
THR
B
308
−35.384
−11.737
−22.317
1.00
19.04
C


ATOM
12854
O
THR
B
308
−35.686
−12.244
−21.233
1.00
18.69
O


ATOM
12856
N
ASP
B
309
−34.264
−11.031
−22.511
1.00
19.36
N


ATOM
12857
CA
ASP
B
309
−33.335
−10.751
−21.421
1.00
19.65
C


ATOM
12859
CB
ASP
B
309
−32.121
−9.983
−21.942
1.00
19.92
C


ATOM
12862
CG
ASP
B
309
−31.171
−9.536
−20.814
1.00
23.51
C


ATOM
12863
OD1
ASP
B
309
−30.520
−10.412
−20.175
1.00
27.27
O


ATOM
12864
OD2
ASP
B
309
−31.076
−8.299
−20.561
1.00
27.40
O


ATOM
12865
C
ASP
B
309
−34.048
−9.977
−20.295
1.00
19.08
C


ATOM
12866
O
ASP
B
309
−33.912
−10.308
−19.124
1.00
18.55
O


ATOM
12868
N
ALA
B
310
−34.836
−8.975
−20.665
1.00
18.93
N


ATOM
12869
CA
ALA
B
310
−35.546
−8.151
−19.688
1.00
19.12
C


ATOM
12871
CB
ALA
B
310
−36.346
−7.062
−20.380
1.00
18.79
C


ATOM
12875
C
ALA
B
310
−36.462
−8.965
−18.784
1.00
19.36
C


ATOM
12876
O
ALA
B
310
−36.560
−8.696
−17.577
1.00
19.58
O


ATOM
12878
N
VAL
B
311
−37.143
−9.942
−19.370
1.00
19.61
N


ATOM
12879
CA
VAL
B
311
−37.996
−10.845
−18.611
1.00
19.68
C


ATOM
12881
CB
VAL
B
311
−38.867
−11.682
−19.555
1.00
19.46
C


ATOM
12883
CG1
VAL
B
311
−39.948
−10.825
−20.123
1.00
18.49
C


ATOM
12887
CG2
VAL
B
311
−39.469
−12.863
−18.843
1.00
19.31
C


ATOM
12891
C
VAL
B
311
−37.161
−11.726
−17.673
1.00
20.46
C


ATOM
12892
O
VAL
B
311
−37.479
−11.843
−16.494
1.00
20.25
O


ATOM
12894
N
GLU
B
312
−36.085
−12.317
−18.189
1.00
21.67
N


ATOM
12895
CA
GLU
B
312
−35.198
−13.168
−17.384
1.00
22.65
C


ATOM
12897
CB
GLU
B
312
−34.056
−13.708
−18.239
1.00
23.03
C


ATOM
12900
CG
GLU
B
312
−34.454
−14.764
−19.261
1.00
24.88
C


ATOM
12903
CD
GLU
B
312
−33.335
−15.077
−20.267
1.00
27.34
C


ATOM
12904
OE1
GLU
B
312
−33.485
−16.057
−21.028
1.00
29.07
O


ATOM
12905
OE2
GLU
B
312
−32.316
−14.342
−20.313
1.00
28.63
O


ATOM
12906
C
GLU
B
312
−34.598
−12.443
−16.167
1.00
23.09
C


ATOM
12907
O
GLU
B
312
−34.506
−13.007
−15.082
1.00
23.04
O


ATOM
12909
N
ARG
B
313
−34.186
−11.199
−16.335
1.00
23.67
N


ATOM
12910
CA
ARG
B
313
−33.590
−10.486
−15.222
1.00
24.60
C


ATOM
12912
CB
ARG
B
313
−32.609
−9.436
−15.735
1.00
25.26
C


ATOM
12915
CG
ARG
B
313
−31.333
−10.030
−16.398
1.00
27.83
C


ATOM
12918
CD
ARG
B
313
−30.289
−8.943
−16.754
1.00
31.87
C


ATOM
12921
NE
ARG
B
313
−30.922
−7.793
−17.430
1.00
35.71
N


ATOM
12923
CZ
ARG
B
313
−31.341
−6.661
−16.837
1.00
38.42
C


ATOM
12924
NH1
ARG
B
313
−31.189
−6.452
−15.522
1.00
39.15
N


ATOM
12927
NH2
ARG
B
313
−31.919
−5.711
−17.576
1.00
39.25
N


ATOM
12930
C
ARG
B
313
−34.638
−9.879
−14.280
1.00
24.67
C


ATOM
12931
O
ARG
B
313
−34.359
−9.645
−13.117
1.00
24.09
O


ATOM
12933
N
TRP
B
314
−35.843
−9.635
−14.781
1.00
25.60
N


ATOM
12934
CA
TRP
B
314
−36.971
−9.180
−13.951
1.00
26.20
C


ATOM
12936
CB
TRP
B
314
−37.488
−10.339
−13.097
1.00
25.90
C


ATOM
12939
CG
TRP
B
314
−38.912
−10.184
−12.662
1.00
23.93
C


ATOM
12940
CD1
TRP
B
314
−39.355
−9.854
−11.424
1.00
22.76
C


ATOM
12942
NE1
TRP
B
314
−40.727
−9.804
−11.412
1.00
21.93
N


ATOM
12944
CE2
TRP
B
314
−41.189
−10.107
−12.659
1.00
20.38
C


ATOM
12945
CD2
TRP
B
314
−40.074
−10.352
−13.473
1.00
21.17
C


ATOM
12946
CE3
TRP
B
314
−40.277
−10.684
−14.811
1.00
20.19
C


ATOM
12948
CZ3
TRP
B
314
−41.549
−10.766
−15.280
1.00
19.79
C


ATOM
12950
CH2
TRP
B
314
−42.640
−10.512
−14.445
1.00
21.00
C


ATOM
12952
CZ2
TRP
B
314
−42.474
−10.179
−13.130
1.00
20.60
C


ATOM
12954
C
TRP
B
314
−36.620
−7.966
−13.076
1.00
27.57
C


ATOM
12955
O
TRP
B
314
−36.890
−7.935
−11.870
1.00
27.56
O


ATOM
12957
N
ASP
B
315
−36.039
−6.959
−13.718
1.00
29.12
N


ATOM
12958
CA
ASP
B
315
−35.452
−5.822
−13.027
1.00
30.46
C


ATOM
12960
CB
ASP
B
315
−33.935
−5.796
−13.315
1.00
30.84
C


ATOM
12963
CG
ASP
B
315
−33.227
−4.551
−12.772
1.00
32.75
C


ATOM
12964
OD1
ASP
B
315
−33.747
−3.904
−11.830
1.00
35.67
O


ATOM
12965
OD2
ASP
B
315
−32.128
−4.222
−13.295
1.00
34.91
O


ATOM
12966
C
ASP
B
315
−36.157
−4.592
−13.557
1.00
31.10
C


ATOM
12967
O
ASP
B
315
−35.888
−4.157
−14.674
1.00
31.35
O


ATOM
12969
N
VAL
B
316
−37.088
−4.044
−12.781
1.00
32.06
N


ATOM
12970
CA
VAL
B
316
−37.864
−2.896
−13.262
1.00
32.65
C


ATOM
12972
CB
VAL
B
316
−39.016
−2.510
−12.334
1.00
32.45
C


ATOM
12974
CG1
VAL
B
316
−39.911
−1.499
−13.012
1.00
32.15
C


ATOM
12978
CG2
VAL
B
316
−38.490
−1.960
−11.040
1.00
32.91
C


ATOM
12982
C
VAL
B
316
−36.964
−1.692
−13.442
1.00
33.43
C


ATOM
12983
O
VAL
B
316
−37.228
−.854
−14.294
1.00
33.74
O


ATOM
12985
N
ASN
B
317
−35.881
−1.628
−12.663
1.00
34.29
N


ATOM
12986
CA
ASN
B
317
−34.937
−.510
−12.730
1.00
34.63
C


ATOM
12988
CB
ASN
B
317
−33.995
−.526
−11.505
1.00
34.50
C


ATOM
12991
CG
ASN
B
317
−34.692
−.084
−10.201
1.00
34.08
C


ATOM
12992
OD1
ASN
B
317
−35.198
1.042
−10.104
1.00
33.72
O


ATOM
12993
ND2
ASN
B
317
−34.690
−.961
−9.193
1.00
31.22
N


ATOM
12996
C
ASN
B
317
−34.124
−.481
−14.029
1.00
35.21
C


ATOM
12997
O
ASN
B
317
−33.196
.296
−14.127
1.00
35.52
O


ATOM
12999
N
ALA
B
318
−34.470
−1.318
−15.013
1.00
35.89
N


ATOM
13000
CA
ALA
B
318
−33.757
−1.382
−16.299
1.00
36.48
C


ATOM
13002
CB
ALA
B
318
−32.727
−2.508
−16.253
1.00
36.49
C


ATOM
13006
C
ALA
B
318
−34.695
−1.525
−17.534
1.00
37.02
C


ATOM
13007
O
ALA
B
318
−34.297
−1.984
−18.616
1.00
36.52
O


ATOM
13009
N
ILE
B
319
−35.952
−1.137
−17.339
1.00
37.76
N


ATOM
13010
CA
ILE
B
319
−36.848
−.733
−18.424
1.00
38.08
C


ATOM
13012
CB
ILE
B
319
−37.926
.247
−17.893
1.00
38.11
C


ATOM
13014
CG1
ILE
B
319
−39.110
−.495
−17.293
1.00
38.08
C


ATOM
13017
CD1
ILE
B
319
−40.068
.451
−16.596
1.00
38.27
C


ATOM
13021
CG2
ILE
B
319
−38.408
1.189
−18.996
1.00
37.84
C


ATOM
13025
C
ILE
B
319
−36.134
.053
−19.516
1.00
38.29
C


ATOM
13026
O
ILE
B
319
−36.192
−.324
−20.677
1.00
38.68
O


ATOM
13028
N
ASN
B
320
−35.460
1.141
−19.125
1.00
38.36
N


ATOM
13029
CA
ASN
B
320
−34.977
2.168
−20.074
1.00
38.17
C


ATOM
13031
CB
ASN
B
320
−34.278
3.331
−19.331
1.00
38.16
C


ATOM
13034
CG
ASN
B
320
−35.258
4.207
−18.514
1.00
37.87
C


ATOM
13035
OD1
ASN
B
320
−36.366
4.542
−18.955
1.00
36.50
O


ATOM
13036
ND2
ASN
B
320
−34.827
4.589
−17.322
1.00
38.12
N


ATOM
13039
C
ASN
B
320
−34.082
1.643
−21.206
1.00
37.78
C


ATOM
13040
O
ASN
B
320
−33.856
2.332
−22.181
1.00
37.76
O


ATOM
13042
N
ASP
B
321
−33.600
.416
−21.083
1.00
37.46
N


ATOM
13043
CA
ASP
B
321
−32.852
−.224
−22.162
1.00
37.25
C


ATOM
13045
CB
ASP
B
321
−32.091
−1.467
−21.629
1.00
37.83
C


ATOM
13048
CG
ASP
B
321
−31.351
−1.207
−20.285
1.00
39.33
C


ATOM
13049
OD1
ASP
B
321
−31.247
−.021
−19.871
1.00
40.72
O


ATOM
13050
OD2
ASP
B
321
−30.887
−2.199
−19.648
1.00
39.62
O


ATOM
13051
C
ASP
B
321
−33.764
−.636
−23.339
1.00
35.99
C


ATOM
13052
O
ASP
B
321
−33.273
−.904
−24.436
1.00
35.87
O


ATOM
13054
N
LEU
B
322
−35.076
−.674
−23.111
1.00
34.66
N


ATOM
13055
CA
LEU
B
322
−36.024
−1.263
−24.061
1.00
33.77
C


ATOM
13057
CB
LEU
B
322
−37.180
−1.961
−23.327
1.00
33.65
C


ATOM
13060
CG
LEU
B
322
−36.994
−3.260
−22.544
1.00
32.41
C


ATOM
13062
CD1
LEU
B
322
−38.223
−3.482
−21.693
1.00
31.54
C


ATOM
13066
CD2
LEU
B
322
−36.769
−4.432
−23.460
1.00
30.73
C


ATOM
13070
C
LEU
B
322
−36.670
−.243
−24.982
1.00
33.32
C


ATOM
13071
O
LEU
B
322
−36.910
.893
−24.570
1.00
33.37
O


ATOM
13073
N
PRO
B
323
−37.005
−.667
−26.218
1.00
32.72
N


ATOM
13074
CA
PRO
B
323
−37.856
.068
−27.126
1.00
32.42
C


ATOM
13076
CB
PRO
B
323
−38.239
−.985
−28.158
1.00
32.28
C


ATOM
13079
CG
PRO
B
323
−37.098
−1.849
−28.231
1.00
32.46
C


ATOM
13082
CD
PRO
B
323
−36.498
−1.891
−26.857
1.00
32.79
C


ATOM
13085
C
PRO
B
323
−39.115
.576
−26.453
1.00
32.50
C


ATOM
13086
O
PRO
B
323
−39.655
−.083
−25.550
1.00
32.46
O


ATOM
13087
N
ASP
B
324
−39.590
1.725
−26.923
1.00
32.44
N


ATOM
13088
CA
ASP
B
324
−40.729
2.387
−26.318
1.00
32.44
C


ATOM
13090
CB
ASP
B
324
−41.059
3.678
−27.071
1.00
32.91
C


ATOM
13093
CG
ASP
B
324
−40.241
4.873
−26.576
1.00
34.17
C


ATOM
13094
OD1
ASP
B
324
−39.507
4.728
−25.562
1.00
35.83
O


ATOM
13095
OD2
ASP
B
324
−40.351
5.960
−27.196
1.00
35.42
O


ATOM
13096
C
ASP
B
324
−41.967
1.504
−26.186
1.00
31.78
C


ATOM
13097
O
ASP
B
324
−42.546
1.440
−25.102
1.00
31.77
O


ATOM
13099
N
TYR
B
325
−42.368
.821
−27.258
1.00
30.99
N


ATOM
13100
CA
TYR
B
325
−43.519
−.086
−27.167
1.00
30.49
C


ATOM
13102
CB
TYR
B
325
−43.941
−.640
−28.536
1.00
30.28
C


ATOM
13105
CG
TYR
B
325
−43.027
−1.688
−29.134
1.00
30.25
C


ATOM
13106
CD1
TYR
B
325
−41.917
−1.332
−29.893
1.00
29.81
C


ATOM
13108
CE1
TYR
B
325
−41.089
−2.300
−30.449
1.00
29.74
C


ATOM
13110
CZ
TYR
B
325
−41.371
−3.643
−30.258
1.00
30.22
C


ATOM
13111
OH
TYR
B
325
−40.565
−4.625
−30.805
1.00
31.28
O


ATOM
13113
CE2
TYR
B
325
−42.466
−4.015
−29.519
1.00
30.23
C


ATOM
13115
CD2
TYR
B
325
−43.288
−3.038
−28.963
1.00
30.49
C


ATOM
13117
C
TYR
B
325
−43.284
−1.221
−26.168
1.00
30.14
C


ATOM
13118
O
TYR
B
325
−44.219
−1.638
−25.483
1.00
30.19
O


ATOM
13120
N
MET
B
326
−42.048
−1.700
−26.060
1.00
29.74
N


ATOM
13121
CA
MET
B
326
−41.755
−2.820
−25.155
1.00
29.82
C


ATOM
13123
CB
MET
B
326
−40.461
−3.541
−25.549
1.00
29.69
C


ATOM
13126
CG
MET
B
326
−40.650
−4.510
−26.686
1.00
28.83
C


ATOM
13129
SD
MET
B
326
−39.123
−5.272
−27.204
1.00
26.99
S


ATOM
13130
CE
MET
B
326
−39.761
−6.499
−28.328
1.00
29.13
C


ATOM
13134
C
MET
B
326
−41.696
−2.402
−23.683
1.00
29.92
C


ATOM
13135
O
MET
B
326
−42.032
−3.204
−22.801
1.00
29.75
O


ATOM
13137
N
LYS
B
327
−41.251
−1.166
−23.429
1.00
29.68
N


ATOM
13138
CA
LYS
B
327
−41.318
−.578
−22.088
1.00
29.46
C


ATOM
13140
CB
LYS
B
327
−41.016
.921
−22.114
1.00
29.93
C


ATOM
13143
CG
LYS
B
327
−39.572
1.305
−21.951
1.00
31.63
C


ATOM
13146
CD
LYS
B
327
−39.410
2.803
−22.249
1.00
34.93
C


ATOM
13149
CE
LYS
B
327
−37.986
3.291
−22.021
1.00
36.86
C


ATOM
13152
NZ
LYS
B
327
−37.642
4.436
−22.917
1.00
37.83
N


ATOM
13156
C
LYS
B
327
−42.708
−.752
−21.531
1.00
28.46
C


ATOM
13157
O
LYS
B
327
−42.879
−1.314
−20.461
1.00
28.54
O


ATOM
13159
N
LEU
B
328
−43.696
−.258
−22.268
1.00
27.39
N


ATOM
13160
CA
LEU
B
328
−45.056
−.216
−21.785
1.00
26.79
C


ATOM
13162
CB
LEU
B
328
−45.946
.519
−22.773
1.00
26.68
C


ATOM
13165
CG
LEU
B
328
−47.265
1.034
−22.212
1.00
26.00
C


ATOM
13167
CD1
LEU
B
328
−47.010
1.964
−21.036
1.00
24.88
C


ATOM
13171
CD2
LEU
B
328
−48.051
1.740
−23.308
1.00
24.55
C


ATOM
13175
C
LEU
B
328
−45.560
−1.628
−21.587
1.00
26.70
C


ATOM
13176
O
LEU
B
328
−46.110
−1.983
−20.543
1.00
27.03
O


ATOM
13178
N
CYS
B
329
−45.341
−2.453
−22.591
1.00
26.39
N


ATOM
13179
CA
CYS
B
329
−45.750
−3.843
−22.524
1.00
26.12
C


ATOM
13181
CB
CYS
B
329
−45.382
−4.526
−23.838
1.00
26.47
C


ATOM
13184
SG
CYS
B
329
−45.857
−6.236
−23.880
1.00
30.60
S


ATOM
13186
C
CYS
B
329
−45.124
−4.568
−21.319
1.00
24.45
C


ATOM
13187
O
CYS
B
329
−45.829
−5.105
−20.486
1.00
23.93
O


ATOM
13189
N
PHE
B
330
−43.801
−4.553
−21.228
1.00
23.30
N


ATOM
13190
CA
PHE
B
330
−43.087
−5.220
−20.141
1.00
22.47
C


ATOM
13192
CB
PHE
B
330
−41.575
−5.019
−20.267
1.00
22.42
C


ATOM
13195
CG
PHE
B
330
−40.800
−5.518
−19.076
1.00
22.37
C


ATOM
13196
CD1
PHE
B
330
−40.502
−6.864
−18.940
1.00
22.47
C


ATOM
13198
CE1
PHE
B
330
−39.799
−7.339
−17.836
1.00
21.64
C


ATOM
13200
CZ
PHE
B
330
−39.379
−6.472
−16.866
1.00
21.50
C


ATOM
13202
CE2
PHE
B
330
−39.659
−5.122
−16.983
1.00
21.92
C


ATOM
13204
CD2
PHE
B
330
−40.374
−4.648
−18.084
1.00
22.29
C


ATOM
13206
C
PHE
B
330
−43.524
−4.739
−18.765
1.00
21.87
C


ATOM
13207
O
PHE
B
330
−43.779
−5.552
−17.875
1.00
21.98
O


ATOM
13209
N
LEU
B
331
−43.594
−3.427
−18.573
1.00
20.88
N


ATOM
13210
CA
LEU
B
331
−43.997
−2.894
−17.276
1.00
20.25
C


ATOM
13212
CB
LEU
B
331
−43.927
−1.354
−17.257
1.00
20.06
C


ATOM
13215
CG
LEU
B
331
−44.124
−.628
−15.912
1.00
19.21
C


ATOM
13217
CD1
LEU
B
331
−43.542
−1.408
−14.737
1.00
18.78
C


ATOM
13221
CD2
LEU
B
331
−43.555
.770
−15.957
1.00
15.75
C


ATOM
13225
C
LEU
B
331
−45.398
−3.408
−16.902
1.00
19.99
C


ATOM
13226
O
LEU
B
331
−45.606
−3.860
−15.771
1.00
19.67
O


ATOM
13228
N
ALA
B
332
−46.336
−3.373
−17.860
1.00
19.56
N


ATOM
13229
CA
ALA
B
332
−47.695
−3.882
−17.629
1.00
19.24
C


ATOM
13231
CB
ALA
B
332
−48.528
−3.802
−18.891
1.00
18.58
C


ATOM
13235
C
ALA
B
332
−47.637
−5.321
−17.108
1.00
19.23
C


ATOM
13236
O
ALA
B
332
−48.235
−5.650
−16.079
1.00
19.18
O


ATOM
13238
N
LEU
B
333
−46.891
−6.162
−17.816
1.00
19.16
N


ATOM
13239
CA
LEU
B
333
−46.727
−7.554
−17.438
1.00
19.08
C


ATOM
13241
CB
LEU
B
333
−45.830
−8.260
−18.462
1.00
18.97
C


ATOM
13244
CG
LEU
B
333
−45.511
−9.758
−18.307
1.00
19.18
C


ATOM
13246
CD1
LEU
B
333
−46.779
−10.611
−18.121
1.00
18.09
C


ATOM
13250
CD2
LEU
B
333
−44.676
−10.262
−19.499
1.00
17.17
C


ATOM
13254
C
LEU
B
333
−46.111
−7.616
−16.042
1.00
19.16
C


ATOM
13255
O
LEU
B
333
−46.629
−8.279
−15.137
1.00
18.97
O


ATOM
13257
N
TYR
B
334
−45.013
−6.887
−15.884
1.00
19.23
N


ATOM
13258
CA
TYR
B
334
−44.212
−6.906
−14.664
1.00
19.33
C


ATOM
13260
CB
TYR
B
334
−43.092
−5.873
−14.785
1.00
19.43
C


ATOM
13263
CG
TYR
B
334
−42.190
−5.748
−13.596
1.00
19.44
C


ATOM
13264
CD1
TYR
B
334
−41.006
−6.464
−13.524
1.00
20.60
C


ATOM
13266
CE1
TYR
B
334
−40.148
−6.347
−12.442
1.00
20.77
C


ATOM
13268
CZ
TYR
B
334
−40.474
−5.503
−11.417
1.00
21.51
C


ATOM
13269
OH
TYR
B
334
−39.628
−5.387
−10.353
1.00
21.42
O


ATOM
13271
CE2
TYR
B
334
−41.646
−4.768
−11.466
1.00
22.15
C


ATOM
13273
CD2
TYR
B
334
−42.495
−4.892
−12.565
1.00
20.55
C


ATOM
13275
C
TYR
B
334
−45.065
−6.614
−13.443
1.00
19.37
C


ATOM
13276
O
TYR
B
334
−44.980
−7.337
−12.441
1.00
19.94
O


ATOM
13278
N
ASN
B
335
−45.888
−5.565
−13.527
1.00
18.83
N


ATOM
13279
CA
ASN
B
335
−46.784
−5.236
−12.443
1.00
18.37
C


ATOM
13281
CB
ASN
B
335
−47.452
−3.896
−12.675
1.00
18.52
C


ATOM
13284
CG
ASN
B
335
−46.493
−2.742
−12.518
1.00
19.27
C


ATOM
13285
OD1
ASN
B
335
−45.421
−2.906
−11.953
1.00
21.25
O


ATOM
13286
ND2
ASN
B
335
−46.872
−1.566
−13.022
1.00
19.24
N


ATOM
13289
C
ASN
B
335
−47.812
−6.333
−12.291
1.00
18.22
C


ATOM
13290
O
ASN
B
335
−47.966
−6.891
−11.207
1.00
18.53
O


ATOM
13292
N
THR
B
336
−48.481
−6.697
−13.379
1.00
17.95
N


ATOM
13293
CA
THR
B
336
−49.525
−7.728
−13.301
1.00
17.61
C


ATOM
13295
CB
THR
B
336
−49.980
−8.204
−14.676
1.00
17.36
C


ATOM
13297
OG1
THR
B
336
−50.249
−7.065
−15.500
1.00
17.02
O


ATOM
13299
CG2
THR
B
336
−51.228
−9.044
−14.551
1.00
16.07
C


ATOM
13303
C
THR
B
336
−49.065
−8.941
−12.501
1.00
17.75
C


ATOM
13304
O
THR
B
336
−49.788
−9.429
−11.621
1.00
18.09
O


ATOM
13306
N
ILE
B
337
−47.859
−9.411
−12.785
1.00
17.69
N


ATOM
13307
CA
ILE
B
337
−47.384
−10.630
−12.165
1.00
17.74
C


ATOM
13309
CB
ILE
B
337
−46.228
−11.245
−12.949
1.00
17.40
C


ATOM
13311
CG1
ILE
B
337
−46.795
−11.865
−14.227
1.00
18.44
C


ATOM
13314
CD1
ILE
B
337
−45.767
−12.192
−15.300
1.00
19.26
C


ATOM
13318
CG2
ILE
B
337
−45.568
−12.320
−12.152
1.00
16.25
C


ATOM
13322
C
ILE
B
337
−47.053
−10.395
−10.699
1.00
18.32
C


ATOM
13323
O
ILE
B
337
−47.497
−11.175
−9.838
1.00
18.06
O


ATOM
13325
N
ASN
B
338
−46.321
−9.311
−10.404
1.00
18.89
N


ATOM
13326
CA
ASN
B
338
−45.985
−8.978
−9.007
1.00
19.49
C


ATOM
13328
CB
ASN
B
338
−45.189
−7.690
−8.917
1.00
19.43
C


ATOM
13331
CG
ASN
B
338
−43.789
−7.836
−9.444
1.00
20.83
C


ATOM
13332
OD1
ASN
B
338
−43.292
−8.954
−9.634
1.00
21.52
O


ATOM
13333
ND2
ASN
B
338
−43.124
−6.697
−9.676
1.00
22.32
N


ATOM
13336
C
ASN
B
338
−47.229
−8.835
−8.146
1.00
20.05
C


ATOM
13337
O
ASN
B
338
−47.182
−9.055
−6.953
1.00
19.81
O


ATOM
13339
N
GLU
B
339
−48.337
−8.459
−8.770
1.00
20.81
N


ATOM
13340
CA
GLU
B
339
−49.589
−8.347
−8.086
1.00
21.81
C


ATOM
13342
CB
GLU
B
339
−50.563
−7.544
−8.933
1.00
22.69
C


ATOM
13345
CG
GLU
B
339
−51.240
−6.422
−8.148
1.00
27.28
C


ATOM
13348
CD
GLU
B
339
−52.571
−5.959
−8.768
1.00
32.79
C


ATOM
13349
OE1
GLU
B
339
−52.761
−4.713
−8.897
1.00
36.20
O


ATOM
13350
OE2
GLU
B
339
−53.416
−6.837
−9.114
1.00
34.35
O


ATOM
13351
C
GLU
B
339
−50.170
−9.732
−7.762
1.00
21.59
C


ATOM
13352
O
GLU
B
339
−50.690
−9.946
−6.666
1.00
21.38
O


ATOM
13354
N
ILE
B
340
−50.094
−10.676
−8.700
1.00
21.48
N


ATOM
13355
CA
ILE
B
340
−50.486
−12.049
−8.384
1.00
21.27
C


ATOM
13357
CB
ILE
B
340
−50.437
−12.990
−9.603
1.00
21.17
C


ATOM
13359
CG1
ILE
B
340
−51.478
−12.589
−10.643
1.00
20.99
C


ATOM
13362
CD1
ILE
B
340
−51.245
−13.202
−12.017
1.00
19.72
C


ATOM
13366
CG2
ILE
B
340
−50.702
−14.444
−9.177
1.00
21.09
C


ATOM
13370
C
ILE
B
340
−49.557
−12.585
−7.278
1.00
21.40
C


ATOM
13371
O
ILE
B
340
−50.023
−13.238
−6.334
1.00
21.74
O


ATOM
13373
N
ALA
B
341
−48.256
−12.301
−7.380
1.00
20.90
N


ATOM
13374
CA
ALA
B
341
−47.312
−12.721
−6.349
1.00
20.62
C


ATOM
13376
CB
ALA
B
341
−45.922
−12.260
−6.694
1.00
20.53
C


ATOM
13380
C
ALA
B
341
−47.720
−12.195
−4.969
1.00
20.59
C


ATOM
13381
O
ALA
B
341
−47.606
−12.901
−3.950
1.00
20.57
O


ATOM
13383
N
TYR
B
342
−48.208
−10.958
−4.949
1.00
20.62
N


ATOM
13384
CA
TYR
B
342
−48.600
−10.303
−3.707
1.00
20.59
C


ATOM
13386
CB
TYR
B
342
−48.790
−8.792
−3.907
1.00
20.05
C


ATOM
13389
CG
TYR
B
342
−49.309
−8.143
−2.674
1.00
18.29
C


ATOM
13390
CD1
TYR
B
342
−48.443
−7.683
−1.698
1.00
17.62
C


ATOM
13392
CE1
TYR
B
342
−48.914
−7.111
−.518
1.00
17.91
C


ATOM
13394
CZ
TYR
B
342
−50.285
−7.005
−.298
1.00
18.18
C


ATOM
13395
OH
TYR
B
342
−50.763
−6.433
.877
1.00
16.00
O


ATOM
13397
CE2
TYR
B
342
−51.169
−7.467
−1.275
1.00
18.10
C


ATOM
13399
CD2
TYR
B
342
−50.670
−8.037
−2.450
1.00
17.67
C


ATOM
13401
C
TYR
B
342
−49.863
−10.928
−3.118
1.00
21.48
C


ATOM
13402
O
TYR
B
342
−49.960
−11.089
−1.913
1.00
20.91
O


ATOM
13404
N
ASP
B
343
−50.832
−11.273
−3.959
1.00
22.93
N


ATOM
13405
CA
ASP
B
343
−52.048
−11.925
−3.470
1.00
24.30
C


ATOM
13407
CB
ASP
B
343
−53.010
−12.282
−4.608
1.00
24.59
C


ATOM
13410
CG
ASP
B
343
−53.577
−11.060
−5.334
1.00
26.66
C


ATOM
13411
OD1
ASP
B
343
−53.619
−9.936
−4.756
1.00
28.24
O


ATOM
13412
OD2
ASP
B
343
−54.004
−11.247
−6.506
1.00
29.56
O


ATOM
13413
C
ASP
B
343
−51.660
−13.208
−2.761
1.00
24.97
C


ATOM
13414
O
ASP
B
343
−52.128
−13.495
−1.658
1.00
24.95
O


ATOM
13416
N
ASN
B
344
−50.792
−13.973
−3.413
1.00
25.90
N


ATOM
13417
CA
ASN
B
344
−50.322
−15.252
−2.884
1.00
26.59
C


ATOM
13419
CB
ASN
B
344
−49.623
−16.043
−3.978
1.00
26.74
C


ATOM
13422
CG
ASN
B
344
−50.594
−16.639
−4.933
1.00
28.21
C


ATOM
13423
OD1
ASN
B
344
−51.080
−17.737
−4.688
1.00
32.10
O


ATOM
13424
ND2
ASN
B
344
−50.923
−15.916
−6.016
1.00
28.40
N


ATOM
13427
C
ASN
B
344
−49.421
−15.146
−1.655
1.00
26.78
C


ATOM
13428
O
ASN
B
344
−49.424
−16.053
−.821
1.00
27.06
O


ATOM
13430
N
LEU
B
345
−48.647
−14.067
−1.533
1.00
26.74
N


ATOM
13431
CA
LEU
B
345
−47.970
−13.793
−.261
1.00
26.59
C


ATOM
13433
CB
LEU
B
345
−47.005
−12.618
−.396
1.00
26.25
C


ATOM
13436
CG
LEU
B
345
−46.046
−12.376
.764
1.00
24.65
C


ATOM
13438
CD1
LEU
B
345
−45.258
−13.621
1.115
1.00
22.50
C


ATOM
13442
CD2
LEU
B
345
−45.119
−11.251
.387
1.00
23.54
C


ATOM
13446
C
LEU
B
345
−49.004
−13.503
.840
1.00
27.00
C


ATOM
13447
O
LEU
B
345
−48.903
−14.014
1.947
1.00
26.93
O


ATOM
13449
N
LYS
B
346
−50.008
−12.697
.518
1.00
27.54
N


ATOM
13450
CA
LYS
B
346
−51.012
−12.300
1.491
1.00
27.96
C


ATOM
13452
CB
LYS
B
346
−51.998
−11.288
.889
1.00
28.13
C


ATOM
13455
CG
LYS
B
346
−52.822
−10.527
1.926
1.00
28.70
C


ATOM
13458
CD
LYS
B
346
−53.781
−9.526
1.281
1.00
29.73
C


ATOM
13461
CE
LYS
B
346
−55.205
−10.063
1.133
1.00
31.17
C


ATOM
13464
NZ
LYS
B
346
−55.839
−9.661
−.171
1.00
32.83
N


ATOM
13468
C
LYS
B
346
−51.788
−13.498
1.984
1.00
28.31
C


ATOM
13469
O
LYS
B
346
−52.074
−13.600
3.177
1.00
28.52
O


ATOM
13471
N
ASP
B
347
−52.149
−14.396
1.075
1.00
28.56
N


ATOM
13472
CA
ASP
B
347
−53.102
−15.441
1.427
1.00
29.11
C


ATOM
13474
CB
ASP
B
347
−54.068
−15.736
.268
1.00
29.50
C


ATOM
13477
CG
ASP
B
347
−54.902
−14.500
−.150
1.00
31.04
C


ATOM
13478
OD1
ASP
B
347
−54.961
−13.500
.616
1.00
31.35
O


ATOM
13479
OD2
ASP
B
347
−55.496
−14.534
−1.261
1.00
33.39
O


ATOM
13480
C
ASP
B
347
−52.403
−16.699
1.891
1.00
28.79
C


ATOM
13481
O
ASP
B
347
−52.824
−17.321
2.854
1.00
29.02
O


ATOM
13483
N
LYS
B
348
−51.326
−17.068
1.222
1.00
28.74
N


ATOM
13484
CA
LYS
B
348
−50.612
−18.293
1.564
1.00
28.84
C


ATOM
13486
CB
LYS
B
348
−50.290
−19.101
.299
1.00
29.24
C


ATOM
13489
CG
LYS
B
348
−51.513
−19.470
−.576
1.00
30.97
C


ATOM
13492
CD
LYS
B
348
−51.051
−20.300
−1.803
1.00
33.66
C


ATOM
13495
CE
LYS
B
348
−52.033
−20.244
−2.989
1.00
34.52
C


ATOM
13498
NZ
LYS
B
348
−53.445
−20.542
−2.607
1.00
35.61
N


ATOM
13502
C
LYS
B
348
−49.330
−18.029
2.358
1.00
27.97
C


ATOM
13503
O
LYS
B
348
−48.763
−18.948
2.925
1.00
27.88
O


ATOM
13505
N
GLY
B
349
−48.874
−16.785
2.403
1.00
27.21
N


ATOM
13506
CA
GLY
B
349
−47.625
−16.473
3.075
1.00
26.87
C


ATOM
13509
C
GLY
B
349
−46.437
−17.227
2.514
1.00
26.71
C


ATOM
13510
O
GLY
B
349
−45.576
−17.669
3.271
1.00
26.65
O


ATOM
13512
N
GLU
B
350
−46.409
−17.390
1.191
1.00
26.61
N


ATOM
13513
CA
GLU
B
350
−45.263
−17.953
.470
1.00
26.43
C


ATOM
13515
CB
GLU
B
350
−45.624
−19.310
−.128
1.00
26.85
C


ATOM
13518
CG
GLU
B
350
−45.685
−20.453
.888
1.00
29.63
C


ATOM
13521
CD
GLU
B
350
−44.304
−21.036
1.227
1.00
34.21
C


ATOM
13522
OE1
GLU
B
350
−43.417
−21.045
.328
1.00
36.93
O


ATOM
13523
OE2
GLU
B
350
−44.106
−21.498
2.384
1.00
35.89
O


ATOM
13524
C
GLU
B
350
−44.900
−16.981
−.637
1.00
25.49
C


ATOM
13525
O
GLU
B
350
−45.774
−16.312
−1.181
1.00
25.35
O


ATOM
13527
N
ASN
B
351
−43.616
−16.866
−.955
1.00
24.74
N


ATOM
13528
CA
ASN
B
351
−43.206
−16.046
−2.090
1.00
24.47
C


ATOM
13530
CB
ASN
B
351
−41.851
−15.369
−1.867
1.00
24.94
C


ATOM
13533
CG
ASN
B
351
−41.428
−14.489
−3.062
1.00
26.98
C


ATOM
13534
OD1
ASN
B
351
−41.976
−14.605
−4.170
1.00
28.96
O


ATOM
13535
ND2
ASN
B
351
−40.457
−13.601
−2.834
1.00
29.55
N


ATOM
13538
C
ASN
B
351
−43.095
−16.918
−3.303
1.00
23.33
C


ATOM
13539
O
ASN
B
351
−42.120
−17.641
−3.429
1.00
23.47
O


ATOM
13541
N
ILE
B
352
−44.060
−16.823
−4.208
1.00
22.09
N


ATOM
13542
CA
ILE
B
352
−44.048
−17.644
−5.410
1.00
21.22
C


ATOM
13544
CB
ILE
B
352
−45.452
−18.213
−5.675
1.00
21.23
C


ATOM
13546
CG1
ILE
B
352
−46.464
−17.080
−5.913
1.00
21.11
C


ATOM
13549
CD1
ILE
B
352
−47.346
−17.305
−7.126
1.00
20.33
C


ATOM
13553
CG2
ILE
B
352
−45.875
−19.116
−4.519
1.00
19.79
C


ATOM
13557
C
ILE
B
352
−43.508
−16.947
−6.689
1.00
20.88
C


ATOM
13558
O
ILE
B
352
−43.486
−17.553
−7.758
1.00
20.76
O


ATOM
13560
N
LEU
B
353
−43.030
−15.705
−6.574
1.00
20.44
N


ATOM
13561
CA
LEU
B
353
−42.603
−14.907
−7.750
1.00
19.84
C


ATOM
13563
CB
LEU
B
353
−42.055
−13.527
−7.325
1.00
19.67
C


ATOM
13566
CG
LEU
B
353
−42.003
−12.339
−8.311
1.00
18.47
C


ATOM
13568
CD1
LEU
B
353
−43.306
−12.083
−9.008
1.00
17.03
C


ATOM
13572
CD2
LEU
B
353
−41.604
−11.064
−7.597
1.00
17.08
C


ATOM
13576
C
LEU
B
353
−41.587
−15.628
−8.633
1.00
19.81
C


ATOM
13577
O
LEU
B
353
−41.720
−15.596
−9.851
1.00
19.55
O


ATOM
13579
N
PRO
B
354
−40.578
−16.299
−8.024
1.00
20.01
N


ATOM
13580
CA
PRO
B
354
−39.571
−17.010
−8.822
1.00
19.73
C


ATOM
13582
CB
PRO
B
354
−38.781
−17.796
−7.779
1.00
19.55
C


ATOM
13585
CG
PRO
B
354
−38.928
−17.037
−6.542
1.00
19.61
C


ATOM
13588
CD
PRO
B
354
−40.289
−16.413
−6.578
1.00
19.90
C


ATOM
13591
C
PRO
B
354
−40.187
−17.971
−9.804
1.00
19.87
C


ATOM
13592
O
PRO
B
354
−39.693
−18.103
−10.918
1.00
20.08
O


ATOM
13593
N
TYR
B
355
−41.264
−18.636
−9.387
1.00
20.14
N


ATOM
13594
CA
TYR
B
355
−41.909
−19.665
−10.216
1.00
20.24
C


ATOM
13596
CB
TYR
B
355
−42.878
−20.520
−9.394
1.00
20.40
C


ATOM
13599
CG
TYR
B
355
−42.189
−21.153
−8.214
1.00
21.82
C


ATOM
13600
CD1
TYR
B
355
−41.119
−21.999
−8.412
1.00
22.86
C


ATOM
13602
CE1
TYR
B
355
−40.457
−22.564
−7.353
1.00
25.07
C


ATOM
13604
CZ
TYR
B
355
−40.850
−22.285
−6.053
1.00
25.58
C


ATOM
13605
OH
TYR
B
355
−40.153
−22.887
−5.020
1.00
27.33
O


ATOM
13607
CE2
TYR
B
355
−41.918
−21.433
−5.815
1.00
23.93
C


ATOM
13609
CD2
TYR
B
355
−42.577
−20.867
−6.900
1.00
23.10
C


ATOM
13611
C
TYR
B
355
−42.625
−19.001
−11.361
1.00
19.67
C


ATOM
13612
O
TYR
B
355
−42.455
−19.405
−12.510
1.00
19.75
O


ATOM
13614
N
LEU
B
356
−43.385
−17.954
−11.030
1.00
18.97
N


ATOM
13615
CA
LEU
B
356
−44.180
−17.205
−12.004
1.00
18.26
C


ATOM
13617
CB
LEU
B
356
−45.039
−16.157
−11.293
1.00
18.03
C


ATOM
13620
CG
LEU
B
356
−46.056
−16.727
−10.312
1.00
18.13
C


ATOM
13622
CD1
LEU
B
356
−46.691
−15.613
−9.500
1.00
19.37
C


ATOM
13626
CD2
LEU
B
356
−47.106
−17.521
−11.046
1.00
17.93
C


ATOM
13630
C
LEU
B
356
−43.298
−16.536
−13.056
1.00
17.76
C


ATOM
13631
O
LEU
B
356
−43.598
−16.567
−14.259
1.00
16.93
O


ATOM
13633
N
THR
B
357
−42.201
−15.949
−12.600
1.00
17.56
N


ATOM
13634
CA
THR
B
357
−41.335
−15.215
−13.501
1.00
17.63
C


ATOM
13636
CB
THR
B
357
−40.438
−14.236
−12.759
1.00
17.69
C


ATOM
13638
OG1
THR
B
357
−39.530
−14.974
−11.934
1.00
18.39
O


ATOM
13640
CG2
THR
B
357
−41.288
−13.242
−11.927
1.00
16.47
C


ATOM
13644
C
THR
B
357
−40.477
−16.145
−14.356
1.00
17.55
C


ATOM
13645
O
THR
B
357
−40.154
−15.793
−15.498
1.00
17.17
O


ATOM
13647
N
LYS
B
358
−40.127
−17.321
−13.824
1.00
17.37
N


ATOM
13648
CA
LYS
B
358
−39.462
−18.337
−14.649
1.00
17.55
C


ATOM
13650
CB
LYS
B
358
−39.030
−19.542
−13.827
1.00
17.87
C


ATOM
13653
CG
LYS
B
358
−38.450
−20.718
−14.640
1.00
18.75
C


ATOM
13656
CD
LYS
B
358
−37.013
−20.497
−15.112
1.00
20.18
C


ATOM
13659
CE
LYS
B
358
−36.361
−21.843
−15.457
1.00
21.49
C


ATOM
13662
NZ
LYS
B
358
−35.189
−21.702
−16.361
1.00
22.89
N


ATOM
13666
C
LYS
B
358
−40.403
−18.777
−15.754
1.00
17.43
C


ATOM
13667
O
LYS
B
358
−40.031
−18.775
−16.926
1.00
17.52
O


ATOM
13669
N
ALA
B
359
−41.633
−19.120
−15.384
1.00
17.42
N


ATOM
13670
CA
ALA
B
359
−42.654
−19.471
−16.359
1.00
17.52
C


ATOM
13672
CB
ALA
B
359
−44.011
−19.434
−15.738
1.00
17.16
C


ATOM
13676
C
ALA
B
359
−42.585
−18.521
−17.531
1.00
18.18
C


ATOM
13677
O
ALA
B
359
−42.513
−18.956
−18.675
1.00
18.50
O


ATOM
13679
N
TRP
B
360
−42.556
−17.223
−17.245
1.00
19.14
N


ATOM
13680
CA
TRP
B
360
−42.526
−16.206
−18.301
1.00
19.77
C


ATOM
13682
CB
TRP
B
360
−42.922
−14.837
−17.746
1.00
20.15
C


ATOM
13685
CG
TRP
B
360
−44.377
−14.653
−17.813
1.00
20.50
C


ATOM
13686
CD1
TRP
B
360
−45.259
−14.729
−16.788
1.00
22.15
C


ATOM
13688
NE1
TRP
B
360
−46.536
−14.533
−17.248
1.00
22.84
N


ATOM
13690
CE2
TRP
B
360
−46.487
−14.348
−18.603
1.00
21.68
C


ATOM
13691
CD2
TRP
B
360
−45.134
−14.418
−18.987
1.00
20.85
C


ATOM
13692
CE3
TRP
B
360
−44.802
−14.253
−20.334
1.00
21.21
C


ATOM
13694
CZ3
TRP
B
360
−45.821
−14.025
−21.245
1.00
20.75
C


ATOM
13696
CH2
TRP
B
360
−47.166
−13.961
−20.827
1.00
21.21
C


ATOM
13698
CZ2
TRP
B
360
−47.515
−14.123
−19.516
1.00
20.97
C


ATOM
13700
C
TRP
B
360
−41.215
−16.110
−19.081
1.00
19.96
C


ATOM
13701
O
TRP
B
360
−41.242
−15.791
−20.271
1.00
19.94
O


ATOM
13703
N
ALA
B
361
−40.089
−16.368
−18.420
1.00
20.14
N


ATOM
13704
CA
ALA
B
361
−38.790
−16.437
−19.102
1.00
20.41
C


ATOM
13706
CB
ALA
B
361
−37.676
−16.581
−18.102
1.00
20.67
C


ATOM
13710
C
ALA
B
361
−38.763
−17.618
−20.041
1.00
20.55
C


ATOM
13711
O
ALA
B
361
−38.330
−17.510
−21.187
1.00
20.76
O


ATOM
13713
N
ASP
B
362
−39.230
−18.749
−19.534
1.00
20.57
N


ATOM
13714
CA
ASP
B
362
−39.360
−19.965
−20.326
1.00
20.97
C


ATOM
13716
CB
ASP
B
362
−39.984
−21.082
−19.470
1.00
21.39
C


ATOM
13719
CG
ASP
B
362
−38.938
−21.902
−18.693
1.00
22.80
C


ATOM
13720
OD1
ASP
B
362
−38.836
−23.109
−18.970
1.00
28.57
O


ATOM
13721
OD2
ASP
B
362
−38.210
−21.384
−17.825
1.00
22.78
O


ATOM
13722
C
ASP
B
362
−40.179
−19.748
−21.618
1.00
20.66
C


ATOM
13723
O
ASP
B
362
−39.740
−20.141
−22.701
1.00
20.69
O


ATOM
13725
N
LEU
B
363
−41.346
−19.110
−21.501
1.00
20.22
N


ATOM
13726
CA
LEU
B
363
−42.245
−18.885
−22.648
1.00
19.82
C


ATOM
13728
CB
LEU
B
363
−43.592
−18.327
−22.164
1.00
19.11
C


ATOM
13731
CG
LEU
B
363
−44.644
−17.945
−23.209
1.00
17.02
C


ATOM
13733
CD1
LEU
B
363
−44.829
−19.033
−24.214
1.00
16.20
C


ATOM
13737
CD2
LEU
B
363
−45.979
−17.618
−22.586
1.00
14.21
C


ATOM
13741
C
LEU
B
363
−41.633
−17.950
−23.706
1.00
20.76
C


ATOM
13742
O
LEU
B
363
−41.656
−18.235
−24.910
1.00
21.00
O


ATOM
13744
N
CYS
B
364
−41.088
−16.826
−23.253
1.00
21.42
N


ATOM
13745
CA
CYS
B
364
−40.427
−15.884
−24.147
1.00
21.67
C


ATOM
13747
CB
CYS
B
364
−39.969
−14.639
−23.376
1.00
21.86
C


ATOM
13750
SG
CYS
B
364
−41.327
−13.638
−22.722
1.00
23.36
S


ATOM
13752
C
CYS
B
364
−39.229
−16.547
−24.835
1.00
21.52
C


ATOM
13753
O
CYS
B
364
−39.010
−16.323
−26.037
1.00
21.67
O


ATOM
13755
N
ASN
B
365
−38.454
−17.352
−24.095
1.00
20.83
N


ATOM
13756
CA
ASN
B
365
−37.306
−18.025
−24.713
1.00
20.65
C


ATOM
13758
CB
ASN
B
365
−36.425
−18.756
−23.691
1.00
20.74
C


ATOM
13761
CG
ASN
B
365
−35.330
−17.865
−23.100
1.00
20.53
C


ATOM
13762
OD1
ASN
B
365
−34.553
−17.229
−23.830
1.00
18.26
O


ATOM
13763
ND2
ASN
B
365
−35.252
−17.841
−21.763
1.00
20.69
N


ATOM
13766
C
ASN
B
365
−37.756
−18.987
−25.811
1.00
20.43
C


ATOM
13767
O
ASN
B
365
−36.998
−19.253
−26.759
1.00
20.11
O


ATOM
13769
N
ALA
B
366
−38.987
−19.492
−25.667
1.00
20.01
N


ATOM
13770
CA
ALA
B
366
−39.618
−20.340
−26.665
1.00
19.70
C


ATOM
13772
CB
ALA
B
366
−40.766
−21.113
−26.045
1.00
19.53
C


ATOM
13776
C
ALA
B
366
−40.099
−19.494
−27.838
1.00
19.74
C


ATOM
13777
O
ALA
B
366
−39.846
−19.824
−28.989
1.00
19.61
O


ATOM
13779
N
PHE
B
367
−40.791
−18.399
−27.553
1.00
19.99
N


ATOM
13780
CA
PHE
B
367
−41.138
−17.446
−28.606
1.00
20.36
C


ATOM
13782
CB
PHE
B
367
−41.805
−16.194
−28.026
1.00
20.62
C


ATOM
13785
CG
PHE
B
367
−43.200
−16.403
−27.537
1.00
21.75
C


ATOM
13786
CD1
PHE
B
367
−44.136
−17.071
−28.315
1.00
23.00
C


ATOM
13788
CE1
PHE
B
367
−45.435
−17.242
−27.874
1.00
23.60
C


ATOM
13790
CZ
PHE
B
367
−45.819
−16.731
−26.648
1.00
23.64
C


ATOM
13792
CE2
PHE
B
367
−44.901
−16.049
−25.871
1.00
23.56
C


ATOM
13794
CD2
PHE
B
367
−43.599
−15.883
−26.316
1.00
22.91
C


ATOM
13796
C
PHE
B
367
−39.906
−16.986
−29.393
1.00
20.19
C


ATOM
13797
O
PHE
B
367
−39.934
−16.903
−30.615
1.00
19.98
O


ATOM
13799
N
LEU
B
368
−38.839
−16.659
−28.675
1.00
20.24
N


ATOM
13800
CA
LEU
B
368
−37.629
−16.161
−29.298
1.00
20.31
C


ATOM
13802
CB
LEU
B
368
−36.569
−15.900
−28.232
1.00
20.27
C


ATOM
13805
CG
LEU
B
368
−35.235
−15.313
−28.694
1.00
20.12
C


ATOM
13807
CD1
LEU
B
368
−35.417
−14.069
−29.592
1.00
20.05
C


ATOM
13811
CD2
LEU
B
368
−34.409
−14.987
−27.465
1.00
18.60
C


ATOM
13815
C
LEU
B
368
−37.122
−17.179
−30.295
1.00
20.53
C


ATOM
13816
O
LEU
B
368
−36.844
−16.844
−31.446
1.00
20.49
O


ATOM
13818
N
GLN
B
369
−37.032
−18.426
−29.832
1.00
20.80
N


ATOM
13819
CA
GLN
B
369
−36.565
−19.557
−30.638
1.00
20.90
C


ATOM
13821
CB
GLN
B
369
−36.614
−20.857
−29.815
1.00
20.98
C


ATOM
13824
CG
GLN
B
369
−36.343
−22.159
−30.576
1.00
20.75
C


ATOM
13827
CD
GLN
B
369
−34.946
−22.238
−31.126
1.00
20.29
C


ATOM
13828
OE1
GLN
B
369
−34.718
−21.984
−32.305
1.00
20.17
O


ATOM
13829
NE2
GLN
B
369
−33.996
−22.581
−30.272
1.00
20.33
N


ATOM
13832
C
GLN
B
369
−37.365
−19.718
−31.910
1.00
21.04
C


ATOM
13833
O
GLN
B
369
−36.803
−20.097
−32.929
1.00
20.82
O


ATOM
13835
N
GLU
B
370
−38.668
−19.453
−31.863
1.00
21.80
N


ATOM
13836
CA
GLU
B
370
−39.480
−19.512
−33.087
1.00
22.73
C


ATOM
13838
CB
GLU
B
370
−40.996
−19.518
−32.801
1.00
23.18
C


ATOM
13841
CG
GLU
B
370
−41.449
−20.608
−31.809
1.00
25.92
C


ATOM
13844
CD
GLU
B
370
−42.927
−21.064
−31.957
1.00
29.74
C


ATOM
13845
OE1
GLU
B
370
−43.840
−20.191
−32.101
1.00
30.13
O


ATOM
13846
OE2
GLU
B
370
−43.156
−22.316
−31.892
1.00
31.94
O


ATOM
13847
C
GLU
B
370
−39.079
−18.358
−34.016
1.00
22.62
C


ATOM
13848
O
GLU
B
370
−38.933
−18.565
−35.221
1.00
22.53
O


ATOM
13850
N
ALA
B
371
−38.864
−17.166
−33.456
1.00
22.65
N


ATOM
13851
CA
ALA
B
371
−38.427
−16.023
−34.254
1.00
22.95
C


ATOM
13853
CB
ALA
B
371
−38.375
−14.753
−33.424
1.00
22.97
C


ATOM
13857
C
ALA
B
371
−37.070
−16.296
−34.891
1.00
23.13
C


ATOM
13858
O
ALA
B
371
−36.900
−16.063
−36.091
1.00
23.38
O


ATOM
13860
N
LYS
B
372
−36.113
−16.804
−34.107
1.00
23.15
N


ATOM
13861
CA
LYS
B
372
−34.776
−17.102
−34.640
1.00
23.10
C


ATOM
13863
CB
LYS
B
372
−33.808
−17.633
−33.575
1.00
22.85
C


ATOM
13866
CG
LYS
B
372
−33.414
−16.616
−32.514
1.00
24.28
C


ATOM
13869
CD
LYS
B
372
−31.973
−16.806
−31.969
1.00
26.34
C


ATOM
13872
CE
LYS
B
372
−31.919
−17.407
−30.544
1.00
27.84
C


ATOM
13875
NZ
LYS
B
372
−31.673
−16.405
−29.458
1.00
27.93
N


ATOM
13879
C
LYS
B
372
−34.887
−18.093
−35.790
1.00
23.16
C


ATOM
13880
O
LYS
B
372
−34.231
−17.920
−36.804
1.00
23.73
O


ATOM
13882
N
TRP
B
373
−35.720
−19.122
−35.658
1.00
23.11
N


ATOM
13883
CA
TRP
B
373
−35.844
−20.094
−36.742
1.00
22.99
C


ATOM
13885
CB
TRP
B
373
−36.746
−21.274
−36.370
1.00
22.71
C


ATOM
13888
CG
TRP
B
373
−36.081
−22.343
−35.562
1.00
20.71
C


ATOM
13889
CD1
TRP
B
373
−34.770
−22.740
−35.623
1.00
19.89
C


ATOM
13891
NE1
TRP
B
373
−34.540
−23.757
−34.734
1.00
19.05
N


ATOM
13893
CE2
TRP
B
373
−35.715
−24.050
−34.090
1.00
18.91
C


ATOM
13894
CD2
TRP
B
373
−36.707
−23.184
−34.599
1.00
18.41
C


ATOM
13895
CE3
TRP
B
373
−38.007
−23.288
−34.105
1.00
15.66
C


ATOM
13897
CZ3
TRP
B
373
−38.271
−24.216
−33.130
1.00
15.41
C


ATOM
13899
CH2
TRP
B
373
−37.276
−25.069
−32.647
1.00
16.33
C


ATOM
13901
CZ2
TRP
B
373
−35.994
−25.005
−33.112
1.00
17.53
C


ATOM
13903
C
TRP
B
373
−36.374
−19.416
−37.991
1.00
23.59
C


ATOM
13904
O
TRP
B
373
−35.878
−19.653
−39.096
1.00
23.60
O


ATOM
13906
N
LEU
B
374
−37.374
−18.559
−37.801
1.00
24.32
N


ATOM
13907
CA
LEU
B
374
−38.008
−17.859
−38.915
1.00
24.66
C


ATOM
13909
CB
LEU
B
374
−39.181
−17.019
−38.428
1.00
24.85
C


ATOM
13912
CG
LEU
B
374
−40.112
−16.449
−39.496
1.00
25.45
C


ATOM
13914
CD1
LEU
B
374
−41.002
−17.535
−40.070
1.00
24.83
C


ATOM
13918
CD2
LEU
B
374
−40.959
−15.293
−38.885
1.00
26.92
C


ATOM
13922
C
LEU
B
374
−37.009
−16.975
−39.618
1.00
24.64
C


ATOM
13923
O
LEU
B
374
−36.934
−16.992
−40.830
1.00
24.33
O


ATOM
13925
N
TYR
B
375
−36.232
−16.220
−38.850
1.00
25.09
N


ATOM
13926
CA
TYR
B
375
−35.283
−15.274
−39.433
1.00
25.71
C


ATOM
13928
CB
TYR
B
375
−34.534
−14.479
−38.349
1.00
25.91
C


ATOM
13931
CG
TYR
B
375
−33.536
−13.468
−38.892
1.00
27.11
C


ATOM
13932
CD1
TYR
B
375
−33.950
−12.200
−39.305
1.00
27.98
C


ATOM
13934
CE1
TYR
B
375
−33.039
−11.271
−39.811
1.00
28.61
C


ATOM
13936
CZ
TYR
B
375
−31.693
−11.605
−39.904
1.00
29.13
C


ATOM
13937
OH
TYR
B
375
−30.790
−10.685
−40.397
1.00
29.83
O


ATOM
13939
CE2
TYR
B
375
−31.256
−12.863
−39.501
1.00
28.79
C


ATOM
13941
CD2
TYR
B
375
−32.177
−13.783
−38.995
1.00
28.03
C


ATOM
13943
C
TYR
B
375
−34.305
−16.036
−40.301
1.00
25.64
C


ATOM
13944
O
TYR
B
375
−34.201
−15.788
−41.503
1.00
25.74
O


ATOM
13946
N
ASN
B
376
−33.635
−17.000
−39.685
1.00
25.73
N


ATOM
13947
CA
ASN
B
376
−32.599
−17.777
−40.346
1.00
25.78
C


ATOM
13949
CB
ASN
B
376
−31.739
−18.497
−39.303
1.00
25.73
C


ATOM
13952
CG
ASN
B
376
−31.155
−17.565
−38.273
1.00
25.13
C


ATOM
13953
OD1
ASN
B
376
−30.491
−16.590
−38.601
1.00
25.53
O


ATOM
13954
ND2
ASN
B
376
−31.389
−17.874
−37.013
1.00
24.64
N


ATOM
13957
C
ASN
B
376
−33.126
−18.824
−41.332
1.00
26.04
C


ATOM
13958
O
ASN
B
376
−32.343
−19.644
−41.807
1.00
26.36
O


ATOM
13960
N
LYS
B
377
−34.428
−18.824
−41.632
1.00
26.16
N


ATOM
13961
CA
LYS
B
377
−35.012
−19.811
−42.552
1.00
26.27
C


ATOM
13963
CB
LYS
B
377
−34.575
−19.538
−44.010
1.00
26.51
C


ATOM
13966
CG
LYS
B
377
−35.470
−18.561
−44.802
1.00
28.01
C


ATOM
13969
CD
LYS
B
377
−34.629
−17.592
−45.667
1.00
29.99
C


ATOM
13972
CE
LYS
B
377
−35.493
−16.605
−46.465
1.00
31.02
C


ATOM
13975
NZ
LYS
B
377
−36.420
−15.819
−45.589
1.00
32.17
N


ATOM
13979
C
LYS
B
377
−34.647
−21.243
−42.150
1.00
25.93
C


ATOM
13980
O
LYS
B
377
−34.380
−22.084
−43.003
1.00
25.82
O


ATOM
13982
N
SER
B
378
−34.637
−21.522
−40.852
1.00
25.65
N


ATOM
13983
CA
SER
B
378
−34.324
−22.860
−40.385
1.00
25.54
C


ATOM
13985
CB
SER
B
378
−34.132
−22.877
−38.882
1.00
25.41
C


ATOM
13988
OG
SER
B
378
−32.905
−22.269
−38.570
1.00
25.67
O


ATOM
13990
C
SER
B
378
−35.412
−23.838
−40.790
1.00
25.62
C


ATOM
13991
O
SER
B
378
−36.495
−23.435
−41.199
1.00
25.70
O


ATOM
13993
N
THR
B
379
−35.108
−25.129
−40.690
1.00
25.66
N


ATOM
13994
CA
THR
B
379
−36.038
−26.175
−41.097
1.00
25.55
C


ATOM
13996
CB
THR
B
379
−35.788
−26.635
−42.546
1.00
25.59
C


ATOM
13998
OG1
THR
B
379
−34.391
−26.899
−42.736
1.00
25.46
O


ATOM
14000
CG2
THR
B
379
−36.255
−25.574
−43.528
1.00
25.88
C


ATOM
14004
C
THR
B
379
−35.886
−27.348
−40.160
1.00
25.45
C


ATOM
14005
O
THR
B
379
−35.372
−28.393
−40.548
1.00
25.70
O


ATOM
14007
N
PRO
B
380
−36.335
−27.179
−38.915
1.00
25.37
N


ATOM
14008
CA
PRO
B
380
−36.144
−28.182
−37.876
1.00
25.22
C


ATOM
14010
CB
PRO
B
380
−36.500
−27.437
−36.583
1.00
25.19
C


ATOM
14013
CG
PRO
B
380
−36.643
−26.021
−36.955
1.00
25.83
C


ATOM
14016
CD
PRO
B
380
−37.025
−25.998
−38.390
1.00
25.71
C


ATOM
14019
C
PRO
B
380
−37.052
−29.379
−38.009
1.00
24.82
C


ATOM
14020
O
PRO
B
380
−38.143
−29.280
−38.575
1.00
24.84
O


ATOM
14021
N
THR
B
381
−36.607
−30.493
−37.436
1.00
24.42
N


ATOM
14022
CA
THR
B
381
−37.386
−31.712
−37.429
1.00
23.99
C


ATOM
14024
CB
THR
B
381
−36.614
−32.873
−36.789
1.00
23.99
C


ATOM
14026
OG1
THR
B
381
−36.339
−32.575
−35.418
1.00
24.02
O


ATOM
14028
CG2
THR
B
381
−35.302
−33.117
−37.524
1.00
23.76
C


ATOM
14032
C
THR
B
381
−38.649
−31.461
−36.636
1.00
23.78
C


ATOM
14033
O
THR
B
381
−38.700
−30.548
−35.813
1.00
23.89
O


ATOM
14035
N
PHE
B
382
−39.672
−32.267
−36.889
1.00
23.53
N


ATOM
14036
CA
PHE
B
382
−40.905
−32.204
−36.105
1.00
22.97
C


ATOM
14038
CB
PHE
B
382
−41.870
−33.305
−36.527
1.00
22.89
C


ATOM
14041
CG
PHE
B
382
−43.079
−33.394
−35.655
1.00
22.26
C


ATOM
14042
CD1
PHE
B
382
−44.189
−32.601
−35.910
1.00
21.65
C


ATOM
14044
CE1
PHE
B
382
−45.300
−32.672
−35.099
1.00
20.87
C


ATOM
14046
CZ
PHE
B
382
−45.310
−33.534
−34.016
1.00
20.36
C


ATOM
14048
CE2
PHE
B
382
−44.204
−34.317
−33.744
1.00
20.52
C


ATOM
14050
CD2
PHE
B
382
−43.098
−34.243
−34.556
1.00
21.17
C


ATOM
14052
C
PHE
B
382
−40.677
−32.329
−34.605
1.00
22.69
C


ATOM
14053
O
PHE
B
382
−41.341
−31.657
−33.834
1.00
22.44
O


ATOM
14055
N
ASP
B
383
−39.764
−33.205
−34.191
1.00
22.52
N


ATOM
14056
CA
ASP
B
383
−39.491
−33.375
−32.765
1.00
22.50
C


ATOM
14058
CB
ASP
B
383
−38.531
−34.525
−32.527
1.00
22.43
C


ATOM
14061
CG
ASP
B
383
−39.174
−35.863
−32.711
1.00
22.71
C


ATOM
14062
OD1
ASP
B
383
−40.359
−35.945
−33.074
1.00
22.89
O


ATOM
14063
OD2
ASP
B
383
−38.470
−36.856
−32.492
1.00
25.17
O


ATOM
14064
C
ASP
B
383
−38.954
−32.116
−32.091
1.00
22.43
C


ATOM
14065
O
ASP
B
383
−39.438
−31.758
−31.014
1.00
22.40
O


ATOM
14067
N
ASP
B
384
−37.966
−31.464
−32.713
1.00
22.37
N


ATOM
14068
CA
ASP
B
384
−37.422
−30.191
−32.213
1.00
22.61
C


ATOM
14070
CB
ASP
B
384
−36.317
−29.648
−33.124
1.00
22.82
C


ATOM
14073
CG
ASP
B
384
−34.963
−30.208
−32.807
1.00
23.40
C


ATOM
14074
OD1
ASP
B
384
−34.847
−30.973
−31.832
1.00
25.33
O


ATOM
14075
OD2
ASP
B
384
−34.009
−29.891
−33.545
1.00
24.26
O


ATOM
14076
C
ASP
B
384
−38.482
−29.113
−32.130
1.00
22.56
C


ATOM
14077
O
ASP
B
384
−38.598
−28.429
−31.108
1.00
22.48
O


ATOM
14079
N
TYR
B
385
−39.227
−28.963
−33.227
1.00
22.36
N


ATOM
14080
CA
TYR
B
385
−40.170
−27.870
−33.402
1.00
22.18
C


ATOM
14082
CB
TYR
B
385
−40.738
−27.867
−34.812
1.00
22.05
C


ATOM
14085
CG
TYR
B
385
−41.818
−26.834
−35.019
1.00
22.60
C


ATOM
14086
CD1
TYR
B
385
−41.493
−25.553
−35.437
1.00
23.64
C


ATOM
14088
CE1
TYR
B
385
−42.462
−24.580
−35.630
1.00
25.00
C


ATOM
14090
CZ
TYR
B
385
−43.788
−24.882
−35.410
1.00
26.86
C


ATOM
14091
OH
TYR
B
385
−44.734
−23.883
−35.622
1.00
28.48
O


ATOM
14093
CE2
TYR
B
385
−44.149
−26.171
−34.980
1.00
25.95
C


ATOM
14095
CD2
TYR
B
385
−43.158
−27.133
−34.790
1.00
23.94
C


ATOM
14097
C
TYR
B
385
−41.319
−27.986
−32.446
1.00
22.13
C


ATOM
14098
O
TYR
B
385
−41.703
−27.017
−31.808
1.00
22.95
O


ATOM
14100
N
PHE
B
386
−41.899
−29.169
−32.380
1.00
21.80
N


ATOM
14101
CA
PHE
B
386
−43.048
−29.394
−31.525
1.00
21.62
C


ATOM
14103
CB
PHE
B
386
−43.624
−30.795
−31.768
1.00
21.73
C


ATOM
14106
CG
PHE
B
386
−44.834
−31.104
−30.952
1.00
21.27
C


ATOM
14107
CD1
PHE
B
386
−46.034
−30.447
−31.196
1.00
22.55
C


ATOM
14109
CE1
PHE
B
386
−47.164
−30.732
−30.448
1.00
23.14
C


ATOM
14111
CZ
PHE
B
386
−47.095
−31.699
−29.446
1.00
23.20
C


ATOM
14113
CE2
PHE
B
386
−45.896
−32.358
−29.208
1.00
21.57
C


ATOM
14115
CD2
PHE
B
386
−44.781
−32.057
−29.959
1.00
20.29
C


ATOM
14117
C
PHE
B
386
−42.630
−29.226
−30.073
1.00
21.34
C


ATOM
14118
O
PHE
B
386
−43.353
−28.639
−29.284
1.00
21.44
O


ATOM
14120
N
GLY
B
387
−41.447
−29.725
−29.736
1.00
20.84
N


ATOM
14121
CA
GLY
B
387
−40.935
−29.618
−28.387
1.00
20.57
C


ATOM
14124
C
GLY
B
387
−40.854
−28.186
−27.916
1.00
20.26
C


ATOM
14125
O
GLY
B
387
−40.930
−27.908
−26.724
1.00
20.86
O


ATOM
14127
N
ASN
B
388
−40.691
−27.268
−28.852
1.00
19.81
N


ATOM
14128
CA
ASN
B
388
−40.758
−25.853
−28.536
1.00
19.51
C


ATOM
14130
CB
ASN
B
388
−39.877
−25.086
−29.508
1.00
19.42
C


ATOM
14133
CG
ASN
B
388
−39.593
−23.700
−29.045
1.00
19.26
C


ATOM
14134
OD1
ASN
B
388
−38.916
−23.513
−28.038
1.00
18.93
O


ATOM
14135
ND2
ASN
B
388
−40.105
−22.709
−29.774
1.00
17.99
N


ATOM
14138
C
ASN
B
388
−42.200
−25.320
−28.604
1.00
19.41
C


ATOM
14139
O
ASN
B
388
−42.591
−24.447
−27.833
1.00
19.58
O


ATOM
14141
N
ALA
B
389
−42.989
−25.848
−29.532
1.00
19.11
N


ATOM
14142
CA
ALA
B
389
−44.313
−25.307
−29.808
1.00
18.87
C


ATOM
14144
CB
ALA
B
389
−44.897
−25.949
−31.060
1.00
18.87
C


ATOM
14148
C
ALA
B
389
−45.264
−25.460
−28.634
1.00
18.51
C


ATOM
14149
O
ALA
B
389
−45.916
−24.511
−28.262
1.00
18.33
O


ATOM
14151
N
TRP
B
390
−45.344
−26.652
−28.054
1.00
18.73
N


ATOM
14152
CA
TRP
B
390
−46.258
−26.887
−26.935
1.00
18.74
C


ATOM
14154
CB
TRP
B
390
−46.389
−28.377
−26.569
1.00
18.75
C


ATOM
14157
CG
TRP
B
390
−45.166
−29.122
−26.019
1.00
18.46
C


ATOM
14158
CD1
TRP
B
390
−44.390
−30.001
−26.705
1.00
18.75
C


ATOM
14160
NE1
TRP
B
390
−43.417
−30.519
−25.897
1.00
17.56
N


ATOM
14162
CE2
TRP
B
390
−43.559
−30.005
−24.642
1.00
16.67
C


ATOM
14163
CD2
TRP
B
390
−44.661
−29.124
−24.674
1.00
17.50
C


ATOM
14164
CE3
TRP
B
390
−45.008
−28.448
−23.505
1.00
18.38
C


ATOM
14166
CZ3
TRP
B
390
−44.251
−28.685
−22.347
1.00
18.34
C


ATOM
14168
CH2
TRP
B
390
−43.164
−29.573
−22.361
1.00
16.75
C


ATOM
14170
CZ2
TRP
B
390
−42.805
−30.234
−23.495
1.00
15.75
C


ATOM
14172
C
TRP
B
390
−45.861
−26.066
−25.727
1.00
18.91
C


ATOM
14173
O
TRP
B
390
−46.707
−25.691
−24.919
1.00
19.02
O


ATOM
14175
N
LYS
B
391
−44.570
−25.787
−25.612
1.00
18.95
N


ATOM
14176
CA
LYS
B
391
−44.085
−24.810
−24.643
1.00
18.85
C


ATOM
14178
CB
LYS
B
391
−42.544
−24.888
−24.508
1.00
19.52
C


ATOM
14181
CG
LYS
B
391
−42.023
−25.373
−23.143
1.00
21.17
C


ATOM
14184
CD
LYS
B
391
−40.516
−25.689
−23.191
1.00
22.87
C


ATOM
14187
CE
LYS
B
391
−40.251
−27.159
−23.508
1.00
23.91
C


ATOM
14190
NZ
LYS
B
391
−38.938
−27.354
−24.191
1.00
25.44
N


ATOM
14194
C
LYS
B
391
−44.537
−23.400
−25.032
1.00
17.57
C


ATOM
14195
O
LYS
B
391
−44.937
−22.629
−24.180
1.00
17.66
O


ATOM
14197
N
SER
B
392
−44.490
−23.068
−26.314
1.00
16.65
N


ATOM
14198
CA
SER
B
392
−44.879
−21.718
−26.764
1.00
16.27
C


ATOM
14200
CB
SER
B
392
−44.241
−21.375
−28.119
1.00
16.22
C


ATOM
14203
OG
SER
B
392
−44.937
−21.969
−29.207
1.00
15.94
O


ATOM
14205
C
SER
B
392
−46.384
−21.504
−26.866
1.00
15.87
C


ATOM
14206
O
SER
B
392
−46.825
−20.395
−27.093
1.00
15.56
O


ATOM
14208
N
SER
B
393
−47.167
−22.568
−26.724
1.00
15.92
N


ATOM
14209
CA
SER
B
393
−48.629
−22.474
−26.718
1.00
15.80
C


ATOM
14211
CB
SER
B
393
−49.240
−23.867
−26.630
1.00
15.78
C


ATOM
14214
OG
SER
B
393
−49.025
−24.426
−25.348
1.00
15.00
O


ATOM
14216
C
SER
B
393
−49.097
−21.646
−25.533
1.00
15.94
C


ATOM
14217
O
SER
B
393
−50.115
−20.948
−25.599
1.00
15.60
O


ATOM
14219
N
SER
B
394
−48.296
−21.740
−24.471
1.00
16.14
N


ATOM
14220
CA
SER
B
394
−48.487
−21.091
−23.177
1.00
16.31
C


ATOM
14222
CB
SER
B
394
−49.062
−19.662
−23.271
1.00
16.18
C


ATOM
14225
OG
SER
B
394
−50.472
−19.649
−23.358
1.00
16.57
O


ATOM
14227
C
SER
B
394
−49.316
−22.006
−22.297
1.00
16.34
C


ATOM
14228
O
SER
B
394
−49.822
−21.595
−21.261
1.00
16.47
O


ATOM
14230
N
GLY
B
395
−49.403
−23.268
−22.702
1.00
16.55
N


ATOM
14231
CA
GLY
B
395
−50.103
−24.282
−21.927
1.00
16.79
C


ATOM
14234
C
GLY
B
395
−49.543
−24.391
−20.534
1.00
16.80
C


ATOM
14235
O
GLY
B
395
−50.222
−24.083
−19.556
1.00
16.74
O


ATOM
14237
N
PRO
B
396
−48.291
−24.824
−20.430
1.00
17.07
N


ATOM
14238
CA
PRO
B
396
−47.698
−24.930
−19.103
1.00
17.18
C


ATOM
14240
CB
PRO
B
396
−46.273
−25.399
−19.386
1.00
17.09
C


ATOM
14243
CG
PRO
B
396
−46.087
−25.237
−20.874
1.00
17.77
C


ATOM
14246
CD
PRO
B
396
−47.424
−25.390
−21.470
1.00
17.20
C


ATOM
14249
C
PRO
B
396
−47.707
−23.615
−18.313
1.00
17.17
C


ATOM
14250
O
PRO
B
396
−47.921
−23.644
−17.089
1.00
17.10
O


ATOM
14251
N
LEU
B
397
−47.499
−22.475
−18.983
1.00
17.04
N


ATOM
14252
CA
LEU
B
397
−47.513
−21.183
−18.261
1.00
16.77
C


ATOM
14254
CB
LEU
B
397
−47.116
−19.969
−19.135
1.00
16.84
C


ATOM
14257
CG
LEU
B
397
−47.145
−18.576
−18.458
1.00
16.91
C


ATOM
14259
CD1
LEU
B
397
−46.577
−18.641
−17.096
1.00
18.36
C


ATOM
14263
CD2
LEU
B
397
−46.373
−17.515
−19.196
1.00
16.88
C


ATOM
14267
C
LEU
B
397
−48.894
−20.988
−17.682
1.00
16.24
C


ATOM
14268
O
LEU
B
397
−49.051
−20.710
−16.494
1.00
16.09
O


ATOM
14270
N
GLN
B
398
−49.902
−21.173
−18.515
1.00
15.72
N


ATOM
14271
CA
GLN
B
398
−51.262
−21.093
−18.024
1.00
15.49
C


ATOM
14273
CB
GLN
B
398
−52.267
−21.387
−19.120
1.00
15.51
C


ATOM
14276
CG
GLN
B
398
−52.371
−20.275
−20.118
1.00
16.12
C


ATOM
14279
CD
GLN
B
398
−53.436
−20.545
−21.114
1.00
17.86
C


ATOM
14280
OE1
GLN
B
398
−54.509
−21.028
−20.757
1.00
20.99
O


ATOM
14281
NE2
GLN
B
398
−53.170
−20.239
−22.374
1.00
18.31
N


ATOM
14284
C
GLN
B
398
−51.471
−22.040
−16.873
1.00
15.03
C


ATOM
14285
O
GLN
B
398
−51.974
−21.638
−15.843
1.00
15.21
O


ATOM
14287
N
LEU
B
399
−51.065
−23.291
−17.022
1.00
14.72
N


ATOM
14288
CA
LEU
B
399
−51.361
−24.254
−15.978
1.00
14.65
C


ATOM
14290
CB
LEU
B
399
−51.201
−25.688
−16.475
1.00
14.44
C


ATOM
14293
CG
LEU
B
399
−52.250
−26.191
−17.478
1.00
14.28
C


ATOM
14295
CD1
LEU
B
399
−51.907
−27.633
−17.846
1.00
15.50
C


ATOM
14299
CD2
LEU
B
399
−53.713
−26.077
−16.986
1.00
10.96
C


ATOM
14303
C
LEU
B
399
−50.554
−23.995
−14.704
1.00
14.88
C


ATOM
14304
O
LEU
B
399
−51.100
−24.161
−13.618
1.00
15.21
O


ATOM
14306
N
ILE
B
400
−49.291
−23.562
−14.810
1.00
14.91
N


ATOM
14307
CA
ILE
B
400
−48.532
−23.174
−13.607
1.00
14.89
C


ATOM
14309
CB
ILE
B
400
−47.158
−22.574
−13.907
1.00
15.19
C


ATOM
14311
CG1
ILE
B
400
−46.189
−23.674
−14.353
1.00
16.76
C


ATOM
14314
CD1
ILE
B
400
−44.777
−23.162
−14.716
1.00
18.19
C


ATOM
14318
CG2
ILE
B
400
−46.603
−21.906
−12.665
1.00
13.65
C


ATOM
14322
C
ILE
B
400
−49.288
−22.137
−12.819
1.00
14.77
C


ATOM
14323
O
ILE
B
400
−49.485
−22.302
−11.632
1.00
15.36
O


ATOM
14325
N
PHE
B
401
−49.717
−21.071
−13.486
1.00
14.59
N


ATOM
14326
CA
PHE
B
401
−50.491
−20.001
−12.844
1.00
14.22
C


ATOM
14328
CB
PHE
B
401
−50.825
−18.900
−13.845
1.00
14.12
C


ATOM
14331
CG
PHE
B
401
−49.803
−17.790
−13.872
1.00
13.83
C


ATOM
14332
CD1
PHE
B
401
−50.012
−16.623
−13.173
1.00
12.77
C


ATOM
14334
CE1
PHE
B
401
−49.074
−15.629
−13.189
1.00
13.09
C


ATOM
14336
CZ
PHE
B
401
−47.906
−15.781
−13.902
1.00
12.96
C


ATOM
14338
CE2
PHE
B
401
−47.684
−16.928
−14.592
1.00
12.86
C


ATOM
14340
CD2
PHE
B
401
−48.622
−17.932
−14.574
1.00
13.30
C


ATOM
14342
C
PHE
B
401
−51.765
−20.478
−12.212
1.00
14.26
C


ATOM
14343
O
PHE
B
401
−52.184
−19.944
−11.207
1.00
13.90
O


ATOM
14345
N
ALA
B
402
−52.377
−21.480
−12.828
1.00
15.09
N


ATOM
14346
CA
ALA
B
402
−53.641
−22.054
−12.364
1.00
15.76
C


ATOM
14348
CB
ALA
B
402
−54.264
−22.899
−13.454
1.00
15.57
C


ATOM
14352
C
ALA
B
402
−53.417
−22.896
−11.129
1.00
16.63
C


ATOM
14353
O
ALA
B
402
−54.284
−22.970
−10.259
1.00
17.00
O


ATOM
14355
N
TYR
B
403
−52.253
−23.544
−11.060
1.00
17.50
N


ATOM
14356
CA
TYR
B
403
−51.885
−24.317
−9.894
1.00
17.86
C


ATOM
14358
CB
TYR
B
403
−50.486
−24.911
−10.038
1.00
17.73
C


ATOM
14361
CG
TYR
B
403
−50.006
−25.576
−8.764
1.00
18.24
C


ATOM
14362
CD1
TYR
B
403
−50.401
−26.867
−8.436
1.00
18.12
C


ATOM
14364
CE1
TYR
B
403
−49.971
−27.476
−7.273
1.00
18.07
C


ATOM
14366
CZ
TYR
B
403
−49.145
−26.790
−6.409
1.00
18.62
C


ATOM
14367
OH
TYR
B
403
−48.727
−27.383
−5.244
1.00
17.70
O


ATOM
14369
CE2
TYR
B
403
−48.748
−25.499
−6.703
1.00
18.93
C


ATOM
14371
CD2
TYR
B
403
−49.177
−24.901
−7.876
1.00
18.88
C


ATOM
14373
C
TYR
B
403
−51.966
−23.461
−8.630
1.00
18.48
C


ATOM
14374
O
TYR
B
403
−52.494
−23.908
−7.616
1.00
18.66
O


ATOM
14376
N
PHE
B
404
−51.468
−22.232
−8.682
1.00
18.91
N


ATOM
14377
CA
PHE
B
404
−51.400
−21.430
−7.468
1.00
19.53
C


ATOM
14379
CB
PHE
B
404
−50.395
−20.325
−7.644
1.00
19.35
C


ATOM
14382
CG
PHE
B
404
−49.014
−20.808
−7.799
1.00
18.79
C


ATOM
14383
CD1
PHE
B
404
−48.311
−21.246
−6.701
1.00
17.67
C


ATOM
14385
CE1
PHE
B
404
−47.001
−21.683
−6.832
1.00
18.44
C


ATOM
14387
CZ
PHE
B
404
−46.384
−21.688
−8.078
1.00
18.44
C


ATOM
14389
CE2
PHE
B
404
−47.083
−21.245
−9.190
1.00
18.79
C


ATOM
14391
CD2
PHE
B
404
−48.396
−20.805
−9.047
1.00
18.90
C


ATOM
14393
C
PHE
B
404
−52.733
−20.817
−7.072
1.00
20.49
C


ATOM
14394
O
PHE
B
404
−52.925
−20.389
−5.924
1.00
19.95
O


ATOM
14396
N
ALA
B
405
−53.636
−20.756
−8.043
1.00
21.92
N


ATOM
14397
CA
ALA
B
405
−54.918
−20.106
−7.872
1.00
23.09
C


ATOM
14399
CB
ALA
B
405
−55.333
−19.426
−9.167
1.00
23.15
C


ATOM
14403
C
ALA
B
405
−55.959
−21.113
−7.446
1.00
24.23
C


ATOM
14404
O
ALA
B
405
−57.003
−20.726
−6.925
1.00
24.33
O


ATOM
14406
N
VAL
B
406
−55.657
−22.399
−7.662
1.00
25.80
N


ATOM
14407
CA
VAL
B
406
−56.569
−23.511
−7.355
1.00
26.88
C


ATOM
14409
CB
VAL
B
406
−56.640
−24.514
−8.512
1.00
26.64
C


ATOM
14411
CG1
VAL
B
406
−57.132
−25.854
−8.012
1.00
27.24
C


ATOM
14415
CG2
VAL
B
406
−57.547
−23.986
−9.593
1.00
26.45
C


ATOM
14419
C
VAL
B
406
−56.161
−24.266
−6.094
1.00
27.91
C


ATOM
14420
O
VAL
B
406
−56.932
−24.345
−5.155
1.00
28.28
O


ATOM
14422
N
VAL
B
407
−54.954
−24.825
−6.087
1.00
29.25
N


ATOM
14423
CA
VAL
B
407
−54.443
−25.573
−4.941
1.00
30.16
C


ATOM
14425
CB
VAL
B
407
−53.128
−26.276
−5.279
1.00
30.04
C


ATOM
14427
CG1
VAL
B
407
−52.482
−26.831
−4.032
1.00
30.42
C


ATOM
14431
CG2
VAL
B
407
−53.378
−27.376
−6.280
1.00
30.14
C


ATOM
14435
C
VAL
B
407
−54.208
−24.646
−3.755
1.00
31.34
C


ATOM
14436
O
VAL
B
407
−53.535
−23.618
−3.876
1.00
31.46
O


ATOM
14438
N
GLN
B
408
−54.753
−25.032
−2.604
1.00
32.63
N


ATOM
14439
CA
GLN
B
408
−54.727
−24.191
−1.417
1.00
33.51
C


ATOM
14441
CB
GLN
B
408
−55.891
−24.572
−.514
1.00
33.84
C


ATOM
14444
CG
GLN
B
408
−56.161
−23.548
.577
1.00
35.49
C


ATOM
14447
CD
GLN
B
408
−57.623
−23.161
.656
1.00
37.61
C


ATOM
14448
OE1
GLN
B
408
−58.519
−24.006
.501
1.00
38.23
O


ATOM
14449
NE2
GLN
B
408
−57.876
−21.874
.893
1.00
38.64
N


ATOM
14452
C
GLN
B
408
−53.399
−24.274
−.647
1.00
33.72
C


ATOM
14453
O
GLN
B
408
−52.852
−23.253
−.204
1.00
33.52
O


ATOM
14455
N
ASN
B
409
−52.889
−25.491
−.490
1.00
33.95
N


ATOM
14456
CA
ASN
B
409
−51.642
−25.707
.226
1.00
34.16
C


ATOM
14458
CB
ASN
B
409
−51.865
−26.665
1.391
1.00
34.25
C


ATOM
14461
CG
ASN
B
409
−52.756
−26.069
2.459
1.00
34.39
C


ATOM
14462
OD1
ASN
B
409
−52.269
−25.577
3.480
1.00
34.40
O


ATOM
14463
ND2
ASN
B
409
−54.070
−26.089
2.222
1.00
34.12
N


ATOM
14466
C
ASN
B
409
−50.582
−26.244
−.709
1.00
34.12
C


ATOM
14467
O
ASN
B
409
−50.578
−27.422
−1.046
1.00
34.41
O


ATOM
14469
N
ILE
B
410
−49.681
−25.369
−1.127
1.00
34.07
N


ATOM
14470
CA
ILE
B
410
−48.699
−25.718
−2.138
1.00
34.08
C


ATOM
14472
CB
ILE
B
410
−48.138
−24.455
−2.840
1.00
34.15
C


ATOM
14474
CG1
ILE
B
410
−47.274
−23.610
−1.891
1.00
34.26
C


ATOM
14477
CD1
ILE
B
410
−47.216
−22.139
−2.249
1.00
34.10
C


ATOM
14481
CG2
ILE
B
410
−49.290
−23.634
−3.404
1.00
34.49
C


ATOM
14485
C
ILE
B
410
−47.586
−26.553
−1.533
1.00
33.93
C


ATOM
14486
O
ILE
B
410
−47.181
−26.317
−.405
1.00
33.80
O


ATOM
14488
N
LYS
B
411
−47.123
−27.546
−2.285
1.00
34.12
N


ATOM
14489
CA
LYS
B
411
−46.012
−28.395
−1.874
1.00
34.45
C


ATOM
14491
CB
LYS
B
411
−46.414
−29.873
−1.907
1.00
34.67
C


ATOM
14494
CG
LYS
B
411
−47.850
−30.130
−1.460
1.00
35.79
C


ATOM
14497
CD
LYS
B
411
−48.102
−31.586
−1.052
1.00
37.60
C


ATOM
14500
CE
LYS
B
411
−49.450
−31.728
−.309
1.00
38.80
C


ATOM
14503
NZ
LYS
B
411
−49.568
−32.992
.488
1.00
39.20
N


ATOM
14507
C
LYS
B
411
−44.843
−28.132
−2.810
1.00
34.33
C


ATOM
14508
O
LYS
B
411
−45.038
−27.956
−4.006
1.00
34.14
O


ATOM
14510
N
LYS
B
412
−43.631
−28.102
−2.265
1.00
34.50
N


ATOM
14511
CA
LYS
B
412
−42.460
−27.688
−3.042
1.00
34.69
C


ATOM
14513
CB
LYS
B
412
−41.242
−27.398
−2.154
1.00
35.13
C


ATOM
14516
CG
LYS
B
412
−41.205
−25.960
−1.613
1.00
36.90
C


ATOM
14519
CD
LYS
B
412
−40.079
−25.749
−.588
1.00
38.57
C


ATOM
14522
CE
LYS
B
412
−40.546
−24.852
.555
1.00
39.55
C


ATOM
14525
NZ
LYS
B
412
−39.491
−24.644
1.587
1.00
40.83
N


ATOM
14529
C
LYS
B
412
−42.075
−28.687
−4.103
1.00
34.13
C


ATOM
14530
O
LYS
B
412
−41.468
−28.308
−5.093
1.00
34.21
O


ATOM
14532
N
GLU
B
413
−42.408
−29.958
−3.910
1.00
33.54
N


ATOM
14533
CA
GLU
B
413
−42.095
−30.949
−4.933
1.00
33.19
C


ATOM
14535
CB
GLU
B
413
−41.886
−32.335
−4.330
1.00
33.44
C


ATOM
14538
CG
GLU
B
413
−43.127
−33.037
−3.807
1.00
34.37
C


ATOM
14541
CD
GLU
B
413
−42.834
−34.490
−3.499
1.00
35.56
C


ATOM
14542
OE1
GLU
B
413
−42.662
−35.264
−4.471
1.00
35.30
O


ATOM
14543
OE2
GLU
B
413
−42.751
−34.847
−2.297
1.00
36.69
O


ATOM
14544
C
GLU
B
413
−43.157
−30.965
−6.029
1.00
32.47
C


ATOM
14545
O
GLU
B
413
−42.846
−31.232
−7.193
1.00
32.28
O


ATOM
14547
N
GLU
B
414
−44.403
−30.676
−5.652
1.00
31.58
N


ATOM
14548
CA
GLU
B
414
−45.482
−30.512
−6.614
1.00
30.99
C


ATOM
14550
CB
GLU
B
414
−46.781
−30.101
−5.927
1.00
30.98
C


ATOM
14553
CG
GLU
B
414
−47.732
−31.245
−5.642
1.00
31.82
C


ATOM
14556
CD
GLU
B
414
−49.100
−30.774
−5.138
1.00
34.29
C


ATOM
14557
OE1
GLU
B
414
−49.258
−29.581
−4.777
1.00
35.53
O


ATOM
14558
OE2
GLU
B
414
−50.036
−31.603
−5.101
1.00
36.44
O


ATOM
14559
C
GLU
B
414
−45.104
−29.455
−7.628
1.00
30.59
C


ATOM
14560
O
GLU
B
414
−45.169
−29.687
−8.828
1.00
30.37
O


ATOM
14562
N
ILE
B
415
−44.684
−28.295
−7.140
1.00
30.49
N


ATOM
14563
CA
ILE
B
415
−44.367
−27.177
−8.028
1.00
30.40
C


ATOM
14565
CB
ILE
B
415
−44.412
−25.797
−7.320
1.00
30.40
C


ATOM
14567
CG1
ILE
B
415
−43.235
−25.589
−6.388
1.00
30.36
C


ATOM
14570
CD1
ILE
B
415
−43.373
−24.313
−5.611
1.00
31.01
C


ATOM
14574
CG2
ILE
B
415
−45.686
−25.644
−6.523
1.00
30.85
C


ATOM
14578
C
ILE
B
415
−43.037
−27.347
−8.730
1.00
30.12
C


ATOM
14579
O
ILE
B
415
−42.870
−26.865
−9.840
1.00
30.42
O


ATOM
14581
N
GLU
B
416
−42.095
−28.030
−8.099
1.00
29.75
N


ATOM
14582
CA
GLU
B
416
−40.799
−28.249
−8.719
1.00
29.70
C


ATOM
14584
CB
GLU
B
416
−39.825
−28.777
−7.690
1.00
30.02
C


ATOM
14587
CG
GLU
B
416
−38.386
−28.426
−7.948
1.00
31.20
C


ATOM
14590
CD
GLU
B
416
−37.523
−28.839
−6.776
1.00
32.91
C


ATOM
14591
OE1
GLU
B
416
−38.008
−28.723
−5.632
1.00
32.37
O


ATOM
14592
OE2
GLU
B
416
−36.377
−29.294
−6.994
1.00
35.31
O


ATOM
14593
C
GLU
B
416
−40.917
−29.222
−9.890
1.00
29.24
C


ATOM
14594
O
GLU
B
416
−40.121
−29.177
−10.835
1.00
28.86
O


ATOM
14596
N
ASN
B
417
−41.915
−30.097
−9.819
1.00
28.81
N


ATOM
14597
CA
ASN
B
417
−42.252
−30.960
−10.941
1.00
28.66
C


ATOM
14599
CB
ASN
B
417
−43.165
−32.105
−10.503
1.00
28.72
C


ATOM
14602
CG
ASN
B
417
−42.379
−33.285
−9.973
1.00
29.78
C


ATOM
14603
OD1
ASN
B
417
−41.887
−34.108
−10.744
1.00
30.39
O


ATOM
14604
ND2
ASN
B
417
−42.223
−33.358
−8.653
1.00
31.45
N


ATOM
14607
C
ASN
B
417
−42.888
−30.183
−12.067
1.00
28.32
C


ATOM
14608
O
ASN
B
417
−42.611
−30.456
−13.232
1.00
27.95
O


ATOM
14610
N
LEU
B
418
−43.740
−29.217
−11.713
1.00
28.23
N


ATOM
14611
CA
LEU
B
418
−44.349
−28.313
−12.697
1.00
27.92
C


ATOM
14613
CB
LEU
B
418
−45.298
−27.320
−12.023
1.00
27.48
C


ATOM
14616
CG
LEU
B
418
−46.636
−27.896
−11.553
1.00
27.02
C


ATOM
14618
CD1
LEU
B
418
−47.393
−26.882
−10.691
1.00
26.70
C


ATOM
14622
CD2
LEU
B
418
−47.499
−28.355
−12.720
1.00
25.90
C


ATOM
14626
C
LEU
B
418
−43.275
−27.567
−13.491
1.00
28.20
C


ATOM
14627
O
LEU
B
418
−43.310
−27.541
−14.733
1.00
27.90
O


ATOM
14629
N
GLN
B
419
−42.308
−26.995
−12.770
1.00
28.43
N


ATOM
14630
CA
GLN
B
419
−41.170
−26.315
−13.395
1.00
28.72
C


ATOM
14632
CB
GLN
B
419
−40.223
−25.746
−12.347
1.00
28.81
C


ATOM
14635
CG
GLN
B
419
−40.592
−24.332
−11.946
1.00
30.14
C


ATOM
14638
CD
GLN
B
419
−39.535
−23.671
−11.092
1.00
32.05
C


ATOM
14639
OE1
GLN
B
419
−39.246
−22.477
−11.257
1.00
33.88
O


ATOM
14640
NE2
GLN
B
419
−38.948
−24.437
−10.170
1.00
31.84
N


ATOM
14643
C
GLN
B
419
−40.390
−27.179
−14.370
1.00
28.83
C


ATOM
14644
O
GLN
B
419
−39.922
−26.668
−15.386
1.00
28.72
O


ATOM
14646
N
LYS
B
420
−40.265
−28.475
−14.067
1.00
29.21
N


ATOM
14647
CA
LYS
B
420
−39.613
−29.451
−14.968
1.00
29.26
C


ATOM
14649
CB
LYS
B
420
−38.924
−30.555
−14.143
1.00
29.42
C


ATOM
14652
CG
LYS
B
420
−37.800
−30.052
−13.207
1.00
30.58
C


ATOM
14655
CD
LYS
B
420
−37.373
−31.113
−12.151
1.00
32.35
C


ATOM
14658
CE
LYS
B
420
−36.572
−30.506
−10.955
1.00
33.05
C


ATOM
14661
NZ
LYS
B
420
−36.520
−31.368
−9.702
1.00
32.62
N


ATOM
14665
C
LYS
B
420
−40.579
−30.057
−16.021
1.00
28.96
C


ATOM
14666
O
LYS
B
420
−40.216
−30.965
−16.753
1.00
28.66
O


ATOM
14668
N
TYR
B
421
−41.804
−29.543
−16.088
1.00
29.01
N


ATOM
14669
CA
TYR
B
421
−42.777
−29.893
−17.134
1.00
29.16
C


ATOM
14671
CB
TYR
B
421
−42.209
−29.602
−18.534
1.00
29.48
C


ATOM
14674
CG
TYR
B
421
−41.968
−28.127
−18.773
1.00
31.04
C


ATOM
14675
CD1
TYR
B
421
−43.018
−27.219
−18.716
1.00
32.54
C


ATOM
14677
CE1
TYR
B
421
−42.815
−25.878
−18.914
1.00
33.35
C


ATOM
14679
CZ
TYR
B
421
−41.556
−25.413
−19.193
1.00
34.56
C


ATOM
14680
OH
TYR
B
421
−41.377
−24.066
−19.384
1.00
37.99
O


ATOM
14682
CE2
TYR
B
421
−40.490
−26.278
−19.267
1.00
33.56
C


ATOM
14684
CD2
TYR
B
421
−40.702
−27.638
−19.051
1.00
32.75
C


ATOM
14686
C
TYR
B
421
−43.345
−31.311
−17.042
1.00
28.60
C


ATOM
14687
O
TYR
B
421
−43.395
−32.046
−18.025
1.00
28.49
O


ATOM
14689
N
HIS
B
422
−43.808
−31.662
−15.846
1.00
28.18
N


ATOM
14690
CA
HIS
B
422
−44.507
−32.919
−15.594
1.00
27.66
C


ATOM
14692
CB
HIS
B
422
−45.082
−32.914
−14.180
1.00
27.73
C


ATOM
14695
CG
HIS
B
422
−45.500
−34.264
−13.685
1.00
28.14
C


ATOM
14696
ND1
HIS
B
422
−44.597
−35.195
−13.217
1.00
29.15
N


ATOM
14698
CE1
HIS
B
422
−45.247
−36.277
−12.829
1.00
29.50
C


ATOM
14700
NE2
HIS
B
422
−46.539
−36.078
−13.022
1.00
28.69
N


ATOM
14702
CD2
HIS
B
422
−46.724
−34.827
−13.552
1.00
27.24
C


ATOM
14704
C
HIS
B
422
−45.650
−33.164
−16.575
1.00
27.29
C


ATOM
14705
O
HIS
B
422
−46.463
−32.269
−16.853
1.00
26.86
O


ATOM
14707
N
ASP
B
423
−45.712
−34.402
−17.060
1.00
26.96
N


ATOM
14708
CA
ASP
B
423
−46.772
−34.883
−17.952
1.00
26.78
C


ATOM
14710
CB
ASP
B
423
−46.792
−36.413
−17.939
1.00
27.01
C


ATOM
14713
CG
ASP
B
423
−45.594
−37.018
−18.646
1.00
28.21
C


ATOM
14714
OD1
ASP
B
423
−45.100
−36.378
−19.602
1.00
31.33
O


ATOM
14715
OD2
ASP
B
423
−45.153
−38.130
−18.262
1.00
28.39
O


ATOM
14716
C
ASP
B
423
−48.187
−34.383
−17.651
1.00
26.21
C


ATOM
14717
O
ASP
B
423
−49.015
−34.336
−18.556
1.00
26.75
O


ATOM
14719
N
ILE
B
424
−48.463
−34.042
−16.391
1.00
25.11
N


ATOM
14720
CA
ILE
B
424
−49.773
−33.553
−15.956
1.00
23.82
C


ATOM
14722
CB
ILE
B
424
−49.798
−33.310
−14.439
1.00
23.67
C


ATOM
14724
CG1
ILE
B
424
−51.196
−32.990
−13.938
1.00
22.97
C


ATOM
14727
CD1
ILE
B
424
−51.220
−32.693
−12.465
1.00
22.06
C


ATOM
14731
CG2
ILE
B
424
−48.859
−32.171
−14.065
1.00
24.35
C


ATOM
14735
C
ILE
B
424
−50.102
−32.265
−16.669
1.00
23.08
C


ATOM
14736
O
ILE
B
424
−51.265
−32.025
−17.023
1.00
22.99
O


ATOM
14738
N
ILE
B
425
−49.082
−31.436
−16.886
1.00
22.22
N


ATOM
14739
CA
ILE
B
425
−49.282
−30.199
−17.633
1.00
21.61
C


ATOM
14741
CB
ILE
B
425
−48.796
−28.959
−16.848
1.00
21.18
C


ATOM
14743
CG1
ILE
B
425
−47.279
−28.853
−16.804
1.00
19.47
C


ATOM
14746
CD1
ILE
B
425
−46.850
−27.581
−16.140
1.00
18.00
C


ATOM
14750
CG2
ILE
B
425
−49.357
−28.977
−15.428
1.00
20.75
C


ATOM
14754
C
ILE
B
425
−48.665
−30.255
−19.033
1.00
21.73
C


ATOM
14755
O
ILE
B
425
−49.100
−29.523
−19.912
1.00
21.75
O


ATOM
14757
N
SER
B
426
−47.686
−31.134
−19.258
1.00
21.74
N


ATOM
14758
CA
SER
B
426
−47.016
−31.174
−20.560
1.00
21.53
C


ATOM
14760
CB
SER
B
426
−45.639
−31.876
−20.516
1.00
21.66
C


ATOM
14763
OG
SER
B
426
−45.724
−33.281
−20.405
1.00
22.19
O


ATOM
14765
C
SER
B
426
−47.922
−31.798
−21.582
1.00
21.19
C


ATOM
14766
O
SER
B
426
−47.950
−31.358
−22.721
1.00
21.37
O


ATOM
14768
N
ARG
B
427
−48.692
−32.800
−21.177
1.00
20.91
N


ATOM
14769
CA
ARG
B
427
−49.532
−33.504
−22.143
1.00
20.78
C


ATOM
14771
CB
ARG
B
427
−50.038
−34.840
−21.616
1.00
20.94
C


ATOM
14774
CG
ARG
B
427
−49.006
−35.887
−21.874
1.00
22.76
C


ATOM
14777
CD
ARG
B
427
−49.158
−37.101
−21.025
1.00
26.75
C


ATOM
14780
NE
ARG
B
427
−47.960
−37.927
−21.185
1.00
29.36
N


ATOM
14782
CZ
ARG
B
427
−47.574
−38.887
−20.352
1.00
30.25
C


ATOM
14783
NH1
ARG
B
427
−48.293
−39.179
−19.266
1.00
30.27
N


ATOM
14786
NH2
ARG
B
427
−46.447
−39.547
−20.611
1.00
31.25
N


ATOM
14789
C
ARG
B
427
−50.646
−32.650
−22.668
1.00
20.07
C


ATOM
14790
O
ARG
B
427
−50.724
−32.466
−23.866
1.00
20.13
O


ATOM
14792
N
PRO
B
428
−51.482
−32.091
−21.786
1.00
19.38
N


ATOM
14793
CA
PRO
B
428
−52.533
−31.202
−22.264
1.00
19.05
C


ATOM
14795
CB
PRO
B
428
−53.046
−30.547
−20.990
1.00
19.13
C


ATOM
14798
CG
PRO
B
428
−52.696
−31.476
−19.918
1.00
19.55
C


ATOM
14801
CD
PRO
B
428
−51.443
−32.161
−20.319
1.00
19.20
C


ATOM
14804
C
PRO
B
428
−52.016
−30.135
−23.227
1.00
18.84
C


ATOM
14805
O
PRO
B
428
−52.688
−29.802
−24.205
1.00
18.73
O


ATOM
14806
N
SER
B
429
−50.828
−29.608
−22.964
1.00
18.57
N


ATOM
14807
CA
SER
B
429
−50.208
−28.679
−23.899
1.00
18.64
C


ATOM
14809
CB
SER
B
429
−48.960
−28.086
−23.291
1.00
18.46
C


ATOM
14812
OG
SER
B
429
−49.141
−27.980
−21.911
1.00
19.08
O


ATOM
14814
C
SER
B
429
−49.877
−29.315
−25.260
1.00
18.52
C


ATOM
14815
O
SER
B
429
−49.886
−28.633
−26.276
1.00
19.02
O


ATOM
14817
N
HIS
B
430
−49.579
−30.606
−25.297
1.00
18.29
N


ATOM
14818
CA
HIS
B
430
−49.457
−31.273
−26.583
1.00
18.25
C


ATOM
14820
CB
HIS
B
430
−49.085
−32.765
−26.467
1.00
18.40
C


ATOM
14823
CG
HIS
B
430
−47.753
−33.024
−25.806
1.00
19.41
C


ATOM
14824
ND1
HIS
B
430
−46.776
−32.055
−25.657
1.00
19.55
N


ATOM
14826
CE1
HIS
B
430
−45.729
−32.576
−25.043
1.00
17.77
C


ATOM
14828
NE2
HIS
B
430
−45.981
−33.849
−24.800
1.00
18.52
N


ATOM
14830
CD2
HIS
B
430
−47.232
−34.160
−25.277
1.00
19.19
C


ATOM
14832
C
HIS
B
430
−50.789
−31.105
−27.306
1.00
17.93
C


ATOM
14833
O
HIS
B
430
−50.816
−30.599
−28.417
1.00
18.54
O


ATOM
14835
N
ILE
B
431
−51.897
−31.479
−26.665
1.00
17.34
N


ATOM
14836
CA
ILE
B
431
−53.217
−31.372
−27.301
1.00
16.58
C


ATOM
14838
CB
ILE
B
431
−54.400
−31.738
−26.375
1.00
16.53
C


ATOM
14840
CG1
ILE
B
431
−54.225
−33.123
−25.728
1.00
16.06
C


ATOM
14843
CD1
ILE
B
431
−54.069
−34.212
−26.698
1.00
16.05
C


ATOM
14847
CG2
ILE
B
431
−55.701
−31.666
−27.149
1.00
15.49
C


ATOM
14851
C
ILE
B
431
−53.431
−29.949
−27.748
1.00
16.35
C


ATOM
14852
O
ILE
B
431
−53.856
−29.712
−28.860
1.00
16.74
O


ATOM
14854
N
PHE
B
432
−53.110
−29.003
−26.881
1.00
16.15
N


ATOM
14855
CA
PHE
B
432
−53.353
−27.577
−27.145
1.00
16.19
C


ATOM
14857
CB
PHE
B
432
−52.811
−26.776
−25.956
1.00
16.36
C


ATOM
14860
CG
PHE
B
432
−53.007
−25.295
−26.043
1.00
16.54
C


ATOM
14861
CD1
PHE
B
432
−53.869
−24.708
−26.946
1.00
16.55
C


ATOM
14863
CE1
PHE
B
432
−54.007
−23.334
−26.974
1.00
17.89
C


ATOM
14865
CZ
PHE
B
432
−53.305
−22.538
−26.080
1.00
17.96
C


ATOM
14867
CE2
PHE
B
432
−52.461
−23.114
−25.169
1.00
17.30
C


ATOM
14869
CD2
PHE
B
432
−52.323
−24.482
−25.151
1.00
17.54
C


ATOM
14871
C
PHE
B
432
−52.726
−27.104
−28.452
1.00
15.93
C


ATOM
14872
O
PHE
B
432
−53.398
−26.525
−29.303
1.00
15.63
O


ATOM
14874
N
ARG
B
433
−51.438
−27.387
−28.594
1.00
15.79
N


ATOM
14875
CA
ARG
B
433
−50.674
−27.044
−29.778
1.00
15.74
C


ATOM
14877
CB
ARG
B
433
−49.196
−27.279
−29.470
1.00
15.68
C


ATOM
14880
CG
ARG
B
433
−48.259
−27.218
−30.665
1.00
16.39
C


ATOM
14883
CD
ARG
B
433
−48.348
−25.919
−31.388
1.00
16.27
C


ATOM
14886
NE
ARG
B
433
−47.969
−24.850
−30.492
1.00
17.34
N


ATOM
14888
CZ
ARG
B
433
−48.210
−23.565
−30.703
1.00
17.89
C


ATOM
14889
NH1
ARG
B
433
−48.834
−23.142
−31.803
1.00
16.79
N


ATOM
14892
NH2
ARG
B
433
−47.805
−22.701
−29.792
1.00
18.53
N


ATOM
14895
C
ARG
B
433
−51.114
−27.852
−31.011
1.00
15.85
C


ATOM
14896
O
ARG
B
433
−51.197
−27.328
−32.120
1.00
15.40
O


ATOM
14898
N
LEU
B
434
−51.381
−29.134
−30.813
1.00
16.18
N


ATOM
14899
CA
LEU
B
434
−51.794
−29.986
−31.902
1.00
16.58
C


ATOM
14901
CB
LEU
B
434
−51.802
−31.453
−31.468
1.00
16.79
C


ATOM
14904
CG
LEU
B
434
−50.408
−32.055
−31.232
1.00
17.49
C


ATOM
14906
CD1
LEU
B
434
−50.486
−33.548
−30.864
1.00
16.63
C


ATOM
14910
CD2
LEU
B
434
−49.511
−31.836
−32.467
1.00
18.00
C


ATOM
14914
C
LEU
B
434
−53.158
−29.559
−32.423
1.00
17.07
C


ATOM
14915
O
LEU
B
434
−53.333
−29.442
−33.633
1.00
17.44
O


ATOM
14917
N
CYS
B
435
−54.118
−29.308
−31.532
1.00
17.23
N


ATOM
14918
CA
CYS
B
435
−55.419
−28.778
−31.947
1.00
17.44
C


ATOM
14920
CB
CYS
B
435
−56.301
−28.529
−30.746
1.00
17.38
C


ATOM
14923
SG
CYS
B
435
−56.825
−29.993
−29.971
1.00
17.90
S


ATOM
14925
C
CYS
B
435
−55.295
−27.460
−32.696
1.00
17.78
C


ATOM
14926
O
CYS
B
435
−55.978
−27.232
−33.701
1.00
17.44
O


ATOM
14928
N
ASN
B
436
−54.442
−26.585
−32.178
1.00
18.31
N


ATOM
14929
CA
ASN
B
436
−54.248
−25.267
−32.769
1.00
19.14
C


ATOM
14931
CB
ASN
B
436
−53.392
−24.394
−31.844
1.00
19.31
C


ATOM
14934
CG
ASN
B
436
−52.984
−23.068
−32.478
1.00
20.07
C


ATOM
14935
OD1
ASN
B
436
−53.456
−22.686
−33.555
1.00
22.72
O


ATOM
14936
ND2
ASN
B
436
−52.099
−22.356
−31.800
1.00
20.90
N


ATOM
14939
C
ASN
B
436
−53.615
−25.345
−34.157
1.00
19.51
C


ATOM
14940
O
ASN
B
436
−54.173
−24.840
−35.131
1.00
19.79
O


ATOM
14942
N
ASP
B
437
−52.446
−25.965
−34.245
1.00
19.85
N


ATOM
14943
CA
ASP
B
437
−51.754
−26.058
−35.517
1.00
20.01
C


ATOM
14945
CB
ASP
B
437
−50.340
−26.638
−35.331
1.00
20.00
C


ATOM
14948
CG
ASP
B
437
−49.397
−25.675
−34.557
1.00
21.23
C


ATOM
14949
OD1
ASP
B
437
−49.902
−24.749
−33.877
1.00
22.90
O


ATOM
14950
OD2
ASP
B
437
−48.149
−25.821
−34.632
1.00
22.18
O


ATOM
14951
C
ASP
B
437
−52.629
−26.838
−36.512
1.00
20.00
C


ATOM
14952
O
ASP
B
437
−52.678
−26.491
−37.695
1.00
20.02
O


ATOM
14954
N
LEU
B
438
−53.372
−27.836
−36.021
1.00
20.02
N


ATOM
14955
CA
LEU
B
438
−54.352
−28.558
−36.858
1.00
20.18
C


ATOM
14957
CB
LEU
B
438
−55.144
−29.593
−36.049
1.00
19.88
C


ATOM
14960
CG
LEU
B
438
−54.699
−31.050
−36.142
1.00
18.97
C


ATOM
14962
CD1
LEU
B
438
−55.537
−31.869
−35.200
1.00
18.86
C


ATOM
14966
CD2
LEU
B
438
−54.818
−31.585
−37.550
1.00
16.23
C


ATOM
14970
C
LEU
B
438
−55.358
−27.644
−37.564
1.00
20.60
C


ATOM
14971
O
LEU
B
438
−55.717
−27.889
−38.712
1.00
20.30
O


ATOM
14973
N
ALA
B
439
−55.825
−26.620
−36.853
1.00
21.39
N


ATOM
14974
CA
ALA
B
439
−56.802
−25.654
−37.377
1.00
21.96
C


ATOM
14976
CB
ALA
B
439
−57.306
−24.741
−36.242
1.00
21.77
C


ATOM
14980
C
ALA
B
439
−56.214
−24.805
−38.494
1.00
22.47
C


ATOM
14981
O
ALA
B
439
−56.850
−24.559
−39.517
1.00
22.13
O


ATOM
14983
N
SER
B
440
−54.983
−24.370
−38.282
1.00
23.40
N


ATOM
14984
CA
SER
B
440
−54.337
−23.428
−39.169
1.00
24.40
C


ATOM
14986
CB
SER
B
440
−53.423
−22.529
−38.342
1.00
24.44
C


ATOM
14989
OG
SER
B
440
−52.994
−23.206
−37.166
1.00
25.19
O


ATOM
14991
C
SER
B
440
−53.544
−24.103
−40.289
1.00
25.15
C


ATOM
14992
O
SER
B
440
−53.093
−23.424
−41.210
1.00
25.24
O


ATOM
14994
N
ALA
B
441
−53.392
−25.428
−40.222
1.00
25.99
N


ATOM
14995
CA
ALA
B
441
−52.488
−26.160
−41.119
1.00
26.53
C


ATOM
14997
CB
ALA
B
441
−52.532
−27.655
−40.830
1.00
26.26
C


ATOM
15001
C
ALA
B
441
−52.726
−25.903
−42.608
1.00
27.31
C


ATOM
15002
O
ALA
B
441
−51.811
−25.489
−43.310
1.00
27.26
O


ATOM
15004
N
SER
B
442
−53.940
−26.134
−43.098
1.00
28.45
N


ATOM
15005
CA
SER
B
442
−54.138
−26.138
−44.545
1.00
29.47
C


ATOM
15007
CB
SER
B
442
−55.477
−26.769
−44.936
1.00
29.40
C


ATOM
15010
OG
SER
B
442
−56.454
−25.784
−45.181
1.00
30.36
O


ATOM
15012
C
SER
B
442
−53.966
−24.731
−45.125
1.00
30.22
C


ATOM
15013
O
SER
B
442
−53.304
−24.562
−46.139
1.00
30.26
O


ATOM
15015
N
ALA
B
443
−54.537
−23.729
−44.467
1.00
31.38
N


ATOM
15016
CA
ALA
B
443
−54.308
−22.333
−44.835
1.00
32.22
C


ATOM
15018
CB
ALA
B
443
−55.057
−21.410
−43.893
1.00
32.28
C


ATOM
15022
C
ALA
B
443
−52.820
−21.992
−44.818
1.00
33.12
C


ATOM
15023
O
ALA
B
443
−52.302
−21.419
−45.766
1.00
33.24
O


ATOM
15025
N
GLU
B
444
−52.136
−22.351
−43.736
1.00
34.43
N


ATOM
15026
CA
GLU
B
444
−50.711
−22.033
−43.582
1.00
35.35
C


ATOM
15028
CB
GLU
B
444
−50.237
−22.271
−42.134
1.00
35.46
C


ATOM
15031
CG
GLU
B
444
−50.757
−21.227
−41.126
1.00
36.42
C


ATOM
15034
CD
GLU
B
444
−50.178
−21.376
−39.708
1.00
37.82
C


ATOM
15035
OE1
GLU
B
444
−49.965
−22.506
−39.224
1.00
37.92
O


ATOM
15036
OE2
GLU
B
444
−49.955
−20.344
−39.049
1.00
40.09
O


ATOM
15037
C
GLU
B
444
−49.840
−22.794
−44.594
1.00
35.87
C


ATOM
15038
O
GLU
B
444
−48.870
−22.239
−45.107
1.00
35.85
O


ATOM
15040
N
ILE
B
445
−50.193
−24.048
−44.883
1.00
36.63
N


ATOM
15041
CA
ILE
B
445
−49.517
−24.837
−45.928
1.00
37.21
C


ATOM
15043
CB
ILE
B
445
−49.854
−26.366
−45.832
1.00
37.15
C


ATOM
15045
CG1
ILE
B
445
−49.181
−26.991
−44.609
1.00
36.85
C


ATOM
15048
CD1
ILE
B
445
−49.867
−28.244
−44.102
1.00
36.62
C


ATOM
15052
CG2
ILE
B
445
−49.416
−27.120
−47.093
1.00
36.66
C


ATOM
15056
C
ILE
B
445
−49.933
−24.289
−47.290
1.00
37.88
C


ATOM
15057
O
ILE
B
445
−50.926
−24.733
−47.874
1.00
38.20
O


ATOM
15059
N
ALA
B
446
−49.175
−23.314
−47.785
1.00
38.56
N


ATOM
15060
CA
ALA
B
446
−49.544
−22.575
−48.992
1.00
39.03
C


ATOM
15062
CB
ALA
B
446
−49.833
−23.522
−50.169
1.00
39.19
C


ATOM
15066
C
ALA
B
446
−50.768
−21.740
−48.685
1.00
39.42
C


ATOM
15067
O
ALA
B
446
−51.866
−22.288
−48.742
1.00
39.26
O


ATOM
15069
N
ARG
B
447
−50.650
−20.443
−48.361
1.00
40.06
N


ATOM
15070
CA
ARG
B
447
−49.426
−19.589
−48.361
1.00
40.54
C


ATOM
15072
CB
ARG
B
447
−49.276
−18.901
−46.986
1.00
40.70
C


ATOM
15075
CG
ARG
B
447
−50.121
−17.634
−46.836
1.00
41.82
C


ATOM
15078
CD
ARG
B
447
−50.292
−17.234
−45.377
1.00
42.99
C


ATOM
15081
NE
ARG
B
447
−51.628
−17.546
−44.864
1.00
44.22
N


ATOM
15083
CZ
ARG
B
447
−51.924
−17.766
−43.580
1.00
46.00
C


ATOM
15084
NH1
ARG
B
447
−50.977
−17.730
−42.633
1.00
46.34
N


ATOM
15087
NH2
ARG
B
447
−53.184
−18.039
−43.236
1.00
46.39
N


ATOM
15090
C
ARG
B
447
−48.095
−20.208
−48.837
1.00
40.35
C


ATOM
15091
O
ARG
B
447
−47.897
−20.403
−50.038
1.00
40.95
O


ATOM
15093
N
GLY
B
448
−47.170
−20.442
−47.918
1.00
39.87
N


ATOM
15094
CA
GLY
B
448
−46.020
−21.306
−48.162
1.00
39.52
C


ATOM
15097
C
GLY
B
448
−45.258
−21.517
−46.865
1.00
39.26
C


ATOM
15098
O
GLY
B
448
−44.071
−21.842
−46.885
1.00
39.15
O


ATOM
15100
N
GLU
B
449
−45.966
−21.344
−45.742
1.00
38.85
N


ATOM
15101
CA
GLU
B
449
−45.365
−21.215
−44.426
1.00
38.45
C


ATOM
15103
CB
GLU
B
449
−46.288
−20.472
−43.444
1.00
38.76
C


ATOM
15106
CG
GLU
B
449
−46.346
−18.943
−43.636
1.00
40.34
C


ATOM
15109
CD
GLU
B
449
−47.486
−18.257
−42.839
1.00
42.09
C


ATOM
15110
OE1
GLU
B
449
−47.897
−18.787
−41.780
1.00
43.23
O


ATOM
15111
OE2
GLU
B
449
−47.972
−17.184
−43.274
1.00
42.13
O


ATOM
15112
C
GLU
B
449
−45.073
−22.600
−43.906
1.00
37.54
C


ATOM
15113
O
GLU
B
449
−45.810
−23.553
−44.167
1.00
37.34
O


ATOM
15115
N
THR
B
450
−43.986
−22.688
−43.159
1.00
36.47
N


ATOM
15116
CA
THR
B
450
−43.459
−23.950
−42.702
1.00
35.65
C


ATOM
15118
CB
THR
B
450
−42.017
−24.105
−43.245
1.00
35.81
C


ATOM
15120
OG1
THR
B
450
−41.593
−25.465
−43.109
1.00
37.67
O


ATOM
15122
CG2
THR
B
450
−41.029
−23.152
−42.538
1.00
36.21
C


ATOM
15126
C
THR
B
450
−43.528
−24.084
−41.166
1.00
34.22
C


ATOM
15127
O
THR
B
450
−43.165
−25.127
−40.619
1.00
34.00
O


ATOM
15129
N
ALA
B
451
−44.024
−23.034
−40.495
1.00
32.69
N


ATOM
15130
CA
ALA
B
451
−44.144
−22.974
−39.026
1.00
31.28
C


ATOM
15132
CB
ALA
B
451
−43.952
−21.546
−38.545
1.00
31.06
C


ATOM
15136
C
ALA
B
451
−45.495
−23.500
−38.547
1.00
29.97
C


ATOM
15137
O
ALA
B
451
−46.337
−22.735
−38.082
1.00
30.08
O


ATOM
15139
N
ASN
B
452
−45.690
−24.809
−38.656
1.00
28.19
N


ATOM
15140
CA
ASN
B
452
−46.935
−25.444
−38.272
1.00
26.73
C


ATOM
15142
CB
ASN
B
452
−47.929
−25.339
−39.422
1.00
26.45
C


ATOM
15145
CG
ASN
B
452
−49.311
−25.832
−39.057
1.00
25.38
C


ATOM
15146
OD1
ASN
B
452
−49.608
−27.004
−39.195
1.00
24.57
O


ATOM
15147
ND2
ASN
B
452
−50.171
−24.928
−38.618
1.00
24.44
N


ATOM
15150
C
ASN
B
452
−46.631
−26.889
−37.952
1.00
26.05
C


ATOM
15151
O
ASN
B
452
−45.930
−27.544
−38.695
1.00
26.21
O


ATOM
15153
N
SER
B
453
−47.131
−27.388
−36.834
1.00
25.30
N


ATOM
15154
CA
SER
B
453
−46.843
−28.760
−36.432
1.00
24.72
C


ATOM
15156
CB
SER
B
453
−47.638
−29.139
−35.174
1.00
24.81
C


ATOM
15159
OG
SER
B
453
−47.143
−28.455
−34.031
1.00
24.36
O


ATOM
15161
C
SER
B
453
−47.111
−29.770
−37.543
1.00
24.12
C


ATOM
15162
O
SER
B
453
−46.325
−30.679
−37.743
1.00
24.33
O


ATOM
15164
N
VAL
B
454
−48.202
−29.606
−38.272
1.00
23.57
N


ATOM
15165
CA
VAL
B
454
−48.546
−30.541
−39.328
1.00
23.27
C


ATOM
15167
CB
VAL
B
454
−50.001
−30.361
−39.795
1.00
23.06
C


ATOM
15169
CG1
VAL
B
454
−50.363
−31.385
−40.829
1.00
22.41
C


ATOM
15173
CG2
VAL
B
454
−50.946
−30.489
−38.622
1.00
22.80
C


ATOM
15177
C
VAL
B
454
−47.581
−30.473
−40.521
1.00
23.83
C


ATOM
15178
O
VAL
B
454
−47.370
−31.493
−41.175
1.00
24.48
O


ATOM
15180
N
SER
B
455
−46.981
−29.309
−40.803
1.00
23.99
N


ATOM
15181
CA
SER
B
455
−45.978
−29.198
−41.881
1.00
24.22
C


ATOM
15183
CB
SER
B
455
−45.577
−27.767
−42.111
1.00
24.02
C


ATOM
15186
OG
SER
B
455
−46.655
−27.098
−42.689
1.00
25.57
O


ATOM
15188
C
SER
B
455
−44.708
−29.949
−41.582
1.00
24.65
C


ATOM
15189
O
SER
B
455
−44.211
−30.700
−42.417
1.00
25.23
O


ATOM
15191
N
CYS
B
456
−44.151
−29.700
−40.405
1.00
24.82
N


ATOM
15192
CA
CYS
B
456
−43.024
−30.462
−39.935
1.00
24.81
C


ATOM
15194
CB
CYS
B
456
−42.765
−30.170
−38.470
1.00
24.92
C


ATOM
15197
SG
CYS
B
456
−42.139
−28.541
−38.190
1.00
25.18
S


ATOM
15199
C
CYS
B
456
−43.304
−31.938
−40.124
1.00
24.84
C


ATOM
15200
O
CYS
B
456
−42.507
−32.634
−40.728
1.00
25.34
O


ATOM
15202
N
TYR
B
457
−44.442
−32.421
−39.644
1.00
24.75
N


ATOM
15203
CA
TYR
B
457
−44.735
−33.838
−39.778
1.00
24.85
C


ATOM
15205
CB
TYR
B
457
−46.069
−34.197
−39.142
1.00
24.64
C


ATOM
15208
CG
TYR
B
457
−46.057
−35.537
−38.451
1.00
24.00
C


ATOM
15209
CD1
TYR
B
457
−45.846
−35.635
−37.083
1.00
24.19
C


ATOM
15211
CE1
TYR
B
457
−45.848
−36.873
−36.436
1.00
23.72
C


ATOM
15213
CZ
TYR
B
457
−46.059
−38.025
−37.170
1.00
23.27
C


ATOM
15214
OH
TYR
B
457
−46.063
−39.244
−36.554
1.00
22.94
O


ATOM
15216
CE2
TYR
B
457
−46.268
−37.961
−38.528
1.00
23.68
C


ATOM
15218
CD2
TYR
B
457
−46.265
−36.711
−39.163
1.00
24.16
C


ATOM
15220
C
TYR
B
457
−44.676
−34.267
−41.251
1.00
25.30
C


ATOM
15221
O
TYR
B
457
−43.965
−35.208
−41.584
1.00
24.94
O


ATOM
15223
N
MET
B
458
−45.388
−33.569
−42.134
1.00
26.02
N


ATOM
15224
CA
MET
B
458
−45.207
−33.775
−43.585
1.00
26.76
C


ATOM
15226
CB
MET
B
458
−45.913
−32.685
−44.407
1.00
26.84
C


ATOM
15229
CG
MET
B
458
−47.419
−32.720
−44.429
1.00
27.32
C


ATOM
15232
SD
MET
B
458
−48.092
−31.238
−45.229
1.00
27.76
S


ATOM
15233
CE
MET
B
458
−47.283
−31.311
−46.830
1.00
26.95
C


ATOM
15237
C
MET
B
458
−43.723
−33.759
−43.996
1.00
27.06
C


ATOM
15238
O
MET
B
458
−43.192
−34.746
−44.485
1.00
26.94
O


ATOM
15240
N
ARG
B
459
−43.069
−32.624
−43.800
1.00
27.58
N


ATOM
15241
CA
ARG
B
459
−41.720
−32.419
−44.289
1.00
28.38
C


ATOM
15243
CB
ARG
B
459
−41.313
−30.965
−44.041
1.00
28.96
C


ATOM
15246
CG
ARG
B
459
−39.862
−30.642
−44.328
1.00
31.93
C


ATOM
15249
CD
ARG
B
459
−39.656
−29.139
−44.480
1.00
36.88
C


ATOM
15252
NE
ARG
B
459
−40.318
−28.349
−43.427
1.00
41.70
N


ATOM
15254
CZ
ARG
B
459
−39.890
−28.241
−42.161
1.00
45.68
C


ATOM
15255
NH1
ARG
B
459
−38.795
−28.899
−41.751
1.00
48.08
N


ATOM
15258
NH2
ARG
B
459
−40.566
−27.489
−41.284
1.00
45.92
N


ATOM
15261
C
ARG
B
459
−40.717
−33.401
−43.672
1.00
28.03
C


ATOM
15262
O
ARG
B
459
−39.829
−33.881
−44.356
1.00
28.13
O


ATOM
15264
N
THR
B
460
−40.882
−33.711
−42.393
1.00
27.88
N


ATOM
15265
CA
THR
B
460
−40.002
−34.635
−41.664
1.00
27.75
C


ATOM
15267
CB
THR
B
460
−40.240
−34.502
−40.134
1.00
27.79
C


ATOM
15269
OG1
THR
B
460
−39.805
−33.209
−39.689
1.00
27.98
O


ATOM
15271
CG2
THR
B
460
−39.519
−35.589
−39.356
1.00
27.46
C


ATOM
15275
C
THR
B
460
−40.171
−36.114
−42.045
1.00
27.72
C


ATOM
15276
O
THR
B
460
−39.211
−36.857
−42.069
1.00
27.60
O


ATOM
15278
N
LYS
B
461
−41.398
−36.538
−42.313
1.00
28.05
N


ATOM
15279
CA
LYS
B
461
−41.701
−37.929
−42.649
1.00
28.25
C


ATOM
15281
CB
LYS
B
461
−42.972
−38.381
−41.914
1.00
28.32
C


ATOM
15284
CG
LYS
B
461
−42.767
−38.763
−40.453
1.00
28.57
C


ATOM
15287
CD
LYS
B
461
−42.569
−40.271
−40.299
1.00
29.79
C


ATOM
15290
CE
LYS
B
461
−41.913
−40.651
−38.970
1.00
30.32
C


ATOM
15293
NZ
LYS
B
461
−42.651
−40.184
−37.749
1.00
30.60
N


ATOM
15297
C
LYS
B
461
−41.876
−38.139
−44.155
1.00
28.50
C


ATOM
15298
O
LYS
B
461
−42.071
−39.269
−44.598
1.00
28.58
O


ATOM
15300
N
GLY
B
462
−41.818
−37.057
−44.933
1.00
28.79
N


ATOM
15301
CA
GLY
B
462
−41.978
−37.114
−46.385
1.00
29.04
C


ATOM
15304
C
GLY
B
462
−43.333
−37.610
−46.853
1.00
29.39
C


ATOM
15305
O
GLY
B
462
−43.410
−38.452
−47.738
1.00
29.53
O


ATOM
15307
N
ILE
B
463
−44.406
−37.081
−46.270
1.00
29.91
N


ATOM
15308
CA
ILE
B
463
−45.764
−37.552
−46.572
1.00
30.27
C


ATOM
15310
CB
ILE
B
463
−46.333
−38.444
−45.433
1.00
30.13
C


ATOM
15312
CG1
ILE
B
463
−46.308
−37.706
−44.092
1.00
30.22
C


ATOM
15315
CD1
ILE
B
463
−46.983
−38.462
−42.965
1.00
30.14
C


ATOM
15319
CG2
ILE
B
463
−45.558
−39.740
−45.340
1.00
29.85
C


ATOM
15323
C
ILE
B
463
−46.775
−36.429
−46.881
1.00
30.75
C


ATOM
15324
O
ILE
B
463
−46.595
−35.270
−46.495
1.00
30.52
O


ATOM
15326
N
SER
B
464
−47.842
−36.822
−47.581
1.00
31.33
N


ATOM
15327
CA
SER
B
464
−48.951
−35.944
−47.952
1.00
31.68
C


ATOM
15329
CB
SER
B
464
−49.948
−36.709
−48.830
1.00
32.02
C


ATOM
15332
OG
SER
B
464
−50.547
−37.793
−48.123
1.00
32.84
O


ATOM
15334
C
SER
B
464
−49.705
−35.396
−46.747
1.00
31.62
C


ATOM
15335
O
SER
B
464
−49.897
−36.097
−45.747
1.00
31.69
O


ATOM
15337
N
GLU
B
465
−50.165
−34.153
−46.874
1.00
31.42
N


ATOM
15338
CA
GLU
B
465
−50.881
−33.466
−45.803
1.00
31.20
C


ATOM
15340
CB
GLU
B
465
−51.465
−32.141
−46.324
1.00
31.33
C


ATOM
15343
CG
GLU
B
465
−52.441
−31.455
−45.359
1.00
31.42
C


ATOM
15346
CD
GLU
B
465
−52.768
−30.006
−45.712
1.00
31.30
C


ATOM
15347
OE1
GLU
B
465
−52.325
−29.490
−46.765
1.00
30.56
O


ATOM
15348
OE2
GLU
B
465
−53.487
−29.380
−44.906
1.00
31.84
O


ATOM
15349
C
GLU
B
465
−51.985
−34.325
−45.202
1.00
30.97
C


ATOM
15350
O
GLU
B
465
−52.175
−34.327
−43.992
1.00
30.72
O


ATOM
15352
N
GLU
B
466
−52.709
−35.052
−46.055
1.00
30.98
N


ATOM
15353
CA
GLU
B
466
−53.865
−35.833
−45.618
1.00
30.65
C


ATOM
15355
CB
GLU
B
466
−54.553
−36.541
−46.788
1.00
30.62
C


ATOM
15358
CG
GLU
B
466
−55.733
−37.401
−46.336
1.00
31.50
C


ATOM
15361
CD
GLU
B
466
−56.353
−38.240
−47.446
1.00
33.14
C


ATOM
15362
OE1
GLU
B
466
−56.512
−37.736
−48.581
1.00
34.40
O


ATOM
15363
OE2
GLU
B
466
−56.699
−39.413
−47.173
1.00
33.21
O


ATOM
15364
C
GLU
B
466
−53.477
−36.851
−44.564
1.00
30.17
C


ATOM
15365
O
GLU
B
466
−54.215
−37.049
−43.606
1.00
30.46
O


ATOM
15367
N
LEU
B
467
−52.333
−37.506
−44.726
1.00
29.59
N


ATOM
15368
CA
LEU
B
467
−51.953
−38.540
−43.769
1.00
29.19
C


ATOM
15370
CB
LEU
B
467
−51.640
−39.869
−44.476
1.00
29.27
C


ATOM
15373
CG
LEU
B
467
−50.356
−40.081
−45.268
1.00
29.84
C


ATOM
15375
CD1
LEU
B
467
−49.341
−40.761
−44.357
1.00
31.06
C


ATOM
15379
CD2
LEU
B
467
−50.589
−40.920
−46.523
1.00
29.47
C


ATOM
15383
C
LEU
B
467
−50.865
−38.092
−42.794
1.00
28.54
C


ATOM
15384
O
LEU
B
467
−50.490
−38.834
−41.894
1.00
28.28
O


ATOM
15386
N
ALA
B
468
−50.388
−36.862
−42.962
1.00
28.04
N


ATOM
15387
CA
ALA
B
468
−49.682
−36.158
−41.893
1.00
27.54
C


ATOM
15389
CB
ALA
B
468
−48.922
−34.970
−42.435
1.00
27.34
C


ATOM
15393
C
ALA
B
468
−50.711
−35.698
−40.865
1.00
27.17
C


ATOM
15394
O
ALA
B
468
−50.482
−35.801
−39.662
1.00
27.40
O


ATOM
15396
N
THR
B
469
−51.841
−35.182
−41.349
1.00
26.57
N


ATOM
15397
CA
THR
B
469
−52.957
−34.795
−40.490
1.00
25.87
C


ATOM
15399
CB
THR
B
469
−54.130
−34.297
−41.328
1.00
25.53
C


ATOM
15401
OG1
THR
B
469
−53.737
−33.111
−42.025
1.00
24.99
O


ATOM
15403
CG2
THR
B
469
−55.316
−33.993
−40.453
1.00
24.62
C


ATOM
15407
C
THR
B
469
−53.412
−35.988
−39.669
1.00
25.95
C


ATOM
15408
O
THR
B
469
−53.551
−35.925
−38.450
1.00
25.80
O


ATOM
15410
N
GLU
B
470
−53.618
−37.091
−40.367
1.00
26.06
N


ATOM
15411
CA
GLU
B
470
−54.043
−38.344
−39.756
1.00
26.10
C


ATOM
15413
CB
GLU
B
470
−54.057
−39.436
−40.848
1.00
26.60
C


ATOM
15416
CG
GLU
B
470
−55.038
−40.587
−40.646
1.00
28.01
C


ATOM
15419
CD
GLU
B
470
−54.553
−41.897
−41.286
1.00
29.05
C


ATOM
15420
OE1
GLU
B
470
−54.298
−41.930
−42.515
1.00
27.96
O


ATOM
15421
OE2
GLU
B
470
−54.435
−42.892
−40.536
1.00
30.42
O


ATOM
15422
C
GLU
B
470
−53.148
−38.746
−38.557
1.00
25.28
C


ATOM
15423
O
GLU
B
470
−53.629
−39.266
−37.556
1.00
25.08
O


ATOM
15425
N
SER
B
471
−51.850
−38.497
−38.654
1.00
24.70
N


ATOM
15426
CA
SER
B
471
−50.930
−38.906
−37.599
1.00
24.26
C


ATOM
15428
CB
SER
B
471
−49.494
−38.842
−38.085
1.00
24.11
C


ATOM
15431
OG
SER
B
471
−49.315
−39.693
−39.186
1.00
23.98
O


ATOM
15433
C
SER
B
471
−51.085
−38.029
−36.377
1.00
23.92
C


ATOM
15434
O
SER
B
471
−51.240
−38.530
−35.270
1.00
24.09
O


ATOM
15436
N
VAL
B
472
−51.032
−36.719
−36.582
1.00
23.48
N


ATOM
15437
CA
VAL
B
472
−51.279
−35.774
−35.510
1.00
23.15
C


ATOM
15439
CB
VAL
B
472
−51.377
−34.332
−36.045
1.00
23.03
C


ATOM
15441
CG1
VAL
B
472
−51.739
−33.347
−34.929
1.00
21.74
C


ATOM
15445
CG2
VAL
B
472
−50.067
−33.945
−36.728
1.00
22.27
C


ATOM
15449
C
VAL
B
472
−52.572
−36.192
−34.830
1.00
23.49
C


ATOM
15450
O
VAL
B
472
−52.663
−36.276
−33.614
1.00
23.29
O


ATOM
15452
N
MET
B
473
−53.573
−36.508
−35.626
1.00
24.34
N


ATOM
15453
CA
MET
B
473
−54.850
−36.901
−35.070
1.00
24.93
C


ATOM
15455
CB
MET
B
473
−55.816
−37.266
−36.190
1.00
25.09
C


ATOM
15458
CG
MET
B
473
−57.191
−36.788
−35.912
1.00
26.94
C


ATOM
15461
SD
MET
B
473
−57.328
−35.035
−36.251
1.00
29.69
S


ATOM
15462
CE
MET
B
473
−58.316
−35.080
−37.775
1.00
29.12
C


ATOM
15466
C
MET
B
473
−54.668
−38.072
−34.100
1.00
25.00
C


ATOM
15467
O
MET
B
473
−55.135
−38.018
−32.963
1.00
24.78
O


ATOM
15469
N
ASN
B
474
−53.965
−39.110
−34.556
1.00
25.30
N


ATOM
15470
CA
ASN
B
474
−53.681
−40.293
−33.738
1.00
25.55
C


ATOM
15472
CB
ASN
B
474
−53.143
−41.439
−34.601
1.00
25.82
C


ATOM
15475
CG
ASN
B
474
−54.243
−42.137
−35.402
1.00
27.62
C


ATOM
15476
OD1
ASN
B
474
−54.238
−42.102
−36.638
1.00
29.33
O


ATOM
15477
ND2
ASN
B
474
−55.196
−42.780
−34.697
1.00
28.58
N


ATOM
15480
C
ASN
B
474
−52.711
−40.034
−32.592
1.00
25.20
C


ATOM
15481
O
ASN
B
474
−52.711
−40.773
−31.621
1.00
25.71
O


ATOM
15483
N
LEU
B
475
−51.883
−38.999
−32.704
1.00
24.65
N


ATOM
15484
CA
LEU
B
475
−50.987
−38.595
−31.614
1.00
24.04
C


ATOM
15486
CB
LEU
B
475
−49.939
−37.626
−32.142
1.00
23.88
C


ATOM
15489
CG
LEU
B
475
−48.837
−37.285
−31.164
1.00
23.70
C


ATOM
15491
CD1
LEU
B
475
−47.936
−38.471
−31.004
1.00
23.97
C


ATOM
15495
CD2
LEU
B
475
−48.088
−36.104
−31.679
1.00
23.99
C


ATOM
15499
C
LEU
B
475
−51.747
−37.940
−30.453
1.00
23.65
C


ATOM
15500
O
LEU
B
475
−51.323
−38.014
−29.299
1.00
23.74
O


ATOM
15502
N
ILE
B
476
−52.858
−37.282
−30.766
1.00
23.15
N


ATOM
15503
CA
ILE
B
476
−53.728
−36.711
−29.742
1.00
22.50
C


ATOM
15505
CB
ILE
B
476
−54.779
−35.755
−30.359
1.00
22.27
C


ATOM
15507
CG1
ILE
B
476
−54.099
−34.508
−30.910
1.00
20.55
C


ATOM
15510
CD1
ILE
B
476
−55.000
−33.671
−31.693
1.00
19.19
C


ATOM
15514
CG2
ILE
B
476
−55.817
−35.363
−29.336
1.00
22.15
C


ATOM
15518
C
ILE
B
476
−54.398
−37.849
−28.985
1.00
22.40
C


ATOM
15519
O
ILE
B
476
−54.316
−37.900
−27.767
1.00
22.32
O


ATOM
15521
N
ASP
B
477
−55.023
−38.777
−29.708
1.00
22.37
N


ATOM
15522
CA
ASP
B
477
−55.616
−39.975
−29.090
1.00
22.55
C


ATOM
15524
CB
ASP
B
477
−55.996
−41.007
−30.151
1.00
22.67
C


ATOM
15527
CG
ASP
B
477
−57.262
−40.636
−30.892
1.00
24.93
C


ATOM
15528
OD1
ASP
B
477
−57.690
−39.458
−30.818
1.00
28.87
O


ATOM
15529
OD2
ASP
B
477
−57.847
−41.523
−31.549
1.00
27.92
O


ATOM
15530
C
ASP
B
477
−54.675
−40.622
−28.083
1.00
22.10
C


ATOM
15531
O
ASP
B
477
−55.048
−40.877
−26.938
1.00
21.83
O


ATOM
15533
N
GLU
B
478
−53.444
−40.867
−28.510
1.00
21.73
N


ATOM
15534
CA
GLU
B
478
−52.472
−41.506
−27.645
1.00
21.69
C


ATOM
15536
CB
GLU
B
478
−51.233
−41.920
−28.437
1.00
21.97
C


ATOM
15539
CG
GLU
B
478
−50.786
−43.360
−28.150
1.00
24.18
C


ATOM
15542
CD
GLU
B
478
−51.793
−44.429
−28.618
1.00
27.03
C


ATOM
15543
OE1
GLU
B
478
−51.599
−45.627
−28.285
1.00
29.15
O


ATOM
15544
OE2
GLU
B
478
−52.770
−44.080
−29.323
1.00
28.37
O


ATOM
15545
C
GLU
B
478
−52.107
−40.608
−26.449
1.00
20.97
C


ATOM
15546
O
GLU
B
478
−51.928
−41.110
−25.316
1.00
21.15
O


ATOM
15548
N
THR
B
479
−52.031
−39.291
−26.687
1.00
19.82
N


ATOM
15549
CA
THR
B
479
−51.787
−38.319
−25.604
1.00
18.59
C


ATOM
15551
CB
THR
B
479
−51.587
−36.897
−26.129
1.00
17.95
C


ATOM
15553
OG1
THR
B
479
−50.291
−36.779
−26.712
1.00
17.43
O


ATOM
15555
CG2
THR
B
479
−51.654
−35.938
−25.009
1.00
17.90
C


ATOM
15559
C
THR
B
479
−52.927
−38.342
−24.580
1.00
18.16
C


ATOM
15560
O
THR
B
479
−52.695
−38.339
−23.383
1.00
18.06
O


ATOM
15562
N
TRP
B
480
−54.160
−38.390
−25.057
1.00
17.77
N


ATOM
15563
CA
TRP
B
480
−55.301
−38.493
−24.169
1.00
17.54
C


ATOM
15565
CB
TRP
B
480
−56.605
−38.464
−24.970
1.00
17.49
C


ATOM
15568
CG
TRP
B
480
−57.239
−37.099
−25.028
1.00
17.53
C


ATOM
15569
CD1
TRP
B
480
−57.150
−36.186
−26.041
1.00
17.04
C


ATOM
15571
NE1
TRP
B
480
−57.857
−35.067
−25.721
1.00
16.57
N


ATOM
15573
CE2
TRP
B
480
−58.423
−35.236
−24.484
1.00
16.41
C


ATOM
15574
CD2
TRP
B
480
−58.055
−36.498
−24.020
1.00
16.68
C


ATOM
15575
CE3
TRP
B
480
−58.507
−36.914
−22.765
1.00
16.92
C


ATOM
15577
CZ3
TRP
B
480
−59.300
−36.076
−22.039
1.00
16.97
C


ATOM
15579
CH2
TRP
B
480
−59.654
−34.823
−22.524
1.00
16.91
C


ATOM
15581
CZ2
TRP
B
480
−59.223
−34.383
−23.742
1.00
16.89
C


ATOM
15583
C
TRP
B
480
−55.232
−39.760
−23.334
1.00
17.75
C


ATOM
15584
O
TRP
B
480
−55.582
−39.747
−22.164
1.00
17.74
O


ATOM
15586
N
LYS
B
481
−54.784
−40.861
−23.933
1.00
18.09
N


ATOM
15587
CA
LYS
B
481
−54.721
−42.124
−23.210
1.00
18.10
C


ATOM
15589
CB
LYS
B
481
−54.277
−43.282
−24.115
1.00
18.01
C


ATOM
15592
CG
LYS
B
481
−55.311
−43.803
−25.114
1.00
16.89
C


ATOM
15595
CD
LYS
B
481
−54.613
−44.663
−26.199
1.00
15.83
C


ATOM
15598
CE
LYS
B
481
−55.587
−45.500
−27.025
1.00
14.74
C


ATOM
15601
NZ
LYS
B
481
−55.119
−45.725
−28.413
1.00
12.34
N


ATOM
15605
C
LYS
B
481
−53.750
−41.970
−22.054
1.00
18.46
C


ATOM
15606
O
LYS
B
481
−53.976
−42.494
−20.969
1.00
18.44
O


ATOM
15608
N
LYS
B
482
−52.662
−41.252
−22.279
1.00
18.80
N


ATOM
15609
CA
LYS
B
482
−51.727
−41.019
−21.192
1.00
19.58
C


ATOM
15611
CB
LYS
B
482
−50.425
−40.424
−21.727
1.00
20.03
C


ATOM
15614
CG
LYS
B
482
−49.499
−41.500
−22.277
1.00
22.03
C


ATOM
15617
CD
LYS
B
482
−48.706
−41.063
−23.506
1.00
24.87
C


ATOM
15620
CE
LYS
B
482
−47.908
−42.268
−24.066
1.00
26.65
C


ATOM
15623
NZ
LYS
B
482
−47.287
−41.996
−25.408
1.00
28.80
N


ATOM
15627
C
LYS
B
482
−52.363
−40.156
−20.096
1.00
19.56
C


ATOM
15628
O
LYS
B
482
−52.367
−40.537
−18.926
1.00
19.19
O


ATOM
15630
N
MET
B
483
−52.922
−39.013
−20.483
1.00
20.00
N


ATOM
15631
CA
MET
B
483
−53.677
−38.165
−19.552
1.00
20.37
C


ATOM
15633
CB
MET
B
483
−54.426
−37.057
−20.295
1.00
20.25
C


ATOM
15636
CG
MET
B
483
−53.529
−35.951
−20.807
1.00
20.52
C


ATOM
15639
SD
MET
B
483
−54.461
−34.483
−21.225
1.00
20.47
S


ATOM
15640
CE
MET
B
483
−55.361
−35.084
−22.649
1.00
22.07
C


ATOM
15644
C
MET
B
483
−54.668
−38.981
−18.741
1.00
20.72
C


ATOM
15645
O
MET
B
483
−54.809
−38.779
−17.543
1.00
20.37
O


ATOM
15647
N
ASN
B
484
−55.345
−39.910
−19.403
1.00
21.57
N


ATOM
15648
CA
ASN
B
484
−56.339
−40.731
−18.744
1.00
22.46
C


ATOM
15650
CB
ASN
B
484
−57.042
−41.631
−19.765
1.00
22.23
C


ATOM
15653
CG
ASN
B
484
−58.118
−40.900
−20.576
1.00
21.61
C


ATOM
15654
OD1
ASN
B
484
−58.412
−39.722
−20.362
1.00
20.25
O


ATOM
15655
ND2
ASN
B
484
−58.720
−41.622
−21.510
1.00
20.80
N


ATOM
15658
C
ASN
B
484
−55.723
−41.555
−17.598
1.00
23.83
C


ATOM
15659
O
ASN
B
484
−56.262
−41.591
−16.484
1.00
23.76
O


ATOM
15661
N
LYS
B
485
−54.585
−42.190
−17.868
1.00
25.69
N


ATOM
15662
CA
LYS
B
485
−53.854
−42.937
−16.843
1.00
27.11
C


ATOM
15664
CB
LYS
B
485
−52.679
−43.712
−17.447
1.00
27.13
C


ATOM
15667
CG
LYS
B
485
−52.073
−44.755
−16.489
1.00
28.85
C


ATOM
15670
CD
LYS
B
485
−51.643
−46.082
−17.187
1.00
31.17
C


ATOM
15673
CE
LYS
B
485
−50.150
−46.132
−17.555
1.00
32.11
C


ATOM
15676
NZ
LYS
B
485
−49.291
−46.282
−16.344
1.00
33.08
N


ATOM
15680
C
LYS
B
485
−53.370
−42.035
−15.701
1.00
28.37
C


ATOM
15681
O
LYS
B
485
−53.288
−42.481
−14.563
1.00
28.59
O


ATOM
15683
N
GLU
B
486
−53.083
−40.766
−15.980
1.00
29.89
N


ATOM
15684
CA
GLU
B
486
−52.683
−39.849
−14.910
1.00
31.09
C


ATOM
15686
CB
GLU
B
486
−52.174
−38.511
−15.470
1.00
31.55
C


ATOM
15689
CG
GLU
B
486
−51.059
−37.851
−14.643
1.00
32.69
C


ATOM
15692
CD
GLU
B
486
−49.785
−38.689
−14.605
1.00
34.51
C


ATOM
15693
OE1
GLU
B
486
−49.104
−38.682
−13.557
1.00
36.35
O


ATOM
15694
OE2
GLU
B
486
−49.476
−39.368
−15.611
1.00
34.55
O


ATOM
15695
C
GLU
B
486
−53.836
−39.615
−13.939
1.00
31.70
C


ATOM
15696
O
GLU
B
486
−53.684
−39.868
−12.750
1.00
31.90
O


ATOM
15698
N
LYS
B
487
−54.983
−39.164
−14.453
1.00
32.71
N


ATOM
15699
CA
LYS
B
487
−56.177
−38.856
−13.629
1.00
33.57
C


ATOM
15701
CB
LYS
B
487
−57.343
−38.402
−14.519
1.00
33.54
C


ATOM
15704
CG
LYS
B
487
−58.703
−38.196
−13.823
1.00
33.45
C


ATOM
15707
CD
LYS
B
487
−58.686
−37.024
−12.866
1.00
33.71
C


ATOM
15710
CE
LYS
B
487
−59.906
−36.996
−11.931
1.00
34.48
C


ATOM
15713
NZ
LYS
B
487
−61.123
−36.378
−12.526
1.00
35.03
N


ATOM
15717
C
LYS
B
487
−56.626
−40.046
−12.802
1.00
34.60
C


ATOM
15718
O
LYS
B
487
−57.126
−39.892
−11.681
1.00
34.55
O


ATOM
15720
N
LEU
B
488
−56.453
−41.233
−13.371
1.00
35.94
N


ATOM
15721
CA
LEU
B
488
−56.878
−42.451
−12.723
1.00
37.00
C


ATOM
15723
CB
LEU
B
488
−57.280
−43.489
−13.778
1.00
36.90
C


ATOM
15726
CG
LEU
B
488
−58.081
−44.714
−13.312
1.00
37.52
C


ATOM
15728
CD1
LEU
B
488
−58.758
−44.536
−11.946
1.00
38.76
C


ATOM
15732
CD2
LEU
B
488
−59.121
−45.084
−14.346
1.00
38.42
C


ATOM
15736
C
LEU
B
488
−55.786
−42.980
−11.803
1.00
38.02
C


ATOM
15737
O
LEU
B
488
−56.063
−43.315
−10.653
1.00
38.19
O


ATOM
15739
N
GLY
B
489
−54.550
−43.016
−12.298
1.00
39.44
N


ATOM
15740
CA
GLY
B
489
−53.441
−43.710
−11.621
1.00
40.62
C


ATOM
15743
C
GLY
B
489
−52.563
−42.837
−10.740
1.00
41.76
C


ATOM
15744
O
GLY
B
489
−51.357
−42.703
−10.988
1.00
41.83
O


ATOM
15746
N
GLY
B
490
−53.183
−42.230
−9.726
1.00
43.10
N


ATOM
15747
CA
GLY
B
490
−52.481
−41.538
−8.636
1.00
43.82
C


ATOM
15750
C
GLY
B
490
−51.131
−40.924
−8.967
1.00
44.31
C


ATOM
15751
O
GLY
B
490
−50.084
−41.560
−8.831
1.00
44.25
O


ATOM
15753
N
SER
B
491
−51.161
−39.672
−9.396
1.00
44.87
N


ATOM
15754
CA
SER
B
491
−49.939
−38.892
−9.575
1.00
45.22
C


ATOM
15756
CB
SER
B
491
−50.224
−37.723
−10.537
1.00
45.34
C


ATOM
15759
OG
SER
B
491
−51.411
−37.032
−10.166
1.00
45.38
O


ATOM
15761
C
SER
B
491
−49.419
−38.389
−8.203
1.00
45.14
C


ATOM
15762
O
SER
B
491
−49.896
−38.824
−7.140
1.00
45.11
O


ATOM
15764
N
LEU
B
492
−48.429
−37.491
−8.227
1.00
44.77
N


ATOM
15765
CA
LEU
B
492
−48.046
−36.752
−7.018
1.00
44.31
C


ATOM
15767
CB
LEU
B
492
−46.726
−35.971
−7.188
1.00
44.55
C


ATOM
15770
CG
LEU
B
492
−45.530
−36.546
−7.972
1.00
45.73
C


ATOM
15772
CD1
LEU
B
492
−44.373
−35.556
−7.859
1.00
46.25
C


ATOM
15776
CD2
LEU
B
492
−45.084
−37.972
−7.530
1.00
46.14
C


ATOM
15780
C
LEU
B
492
−49.151
−35.760
−6.703
1.00
43.30
C


ATOM
15781
O
LEU
B
492
−49.315
−35.370
−5.555
1.00
43.49
O


ATOM
15783
N
PHE
B
493
−49.903
−35.367
−7.735
1.00
42.05
N


ATOM
15784
CA
PHE
B
493
−50.822
−34.239
−7.668
1.00
41.03
C


ATOM
15786
CB
PHE
B
493
−50.913
−33.549
−9.028
1.00
40.81
C


ATOM
15789
CG
PHE
B
493
−49.696
−32.771
−9.408
1.00
38.96
C


ATOM
15790
CD1
PHE
B
493
−49.651
−31.408
−9.233
1.00
37.06
C


ATOM
15792
CE1
PHE
B
493
−48.543
−30.697
−9.603
1.00
36.41
C


ATOM
15794
CZ
PHE
B
493
−47.471
−31.341
−10.158
1.00
35.87
C


ATOM
15796
CE2
PHE
B
493
−47.507
−32.690
−10.343
1.00
36.39
C


ATOM
15798
CD2
PHE
B
493
−48.612
−33.400
−9.977
1.00
37.36
C


ATOM
15800
C
PHE
B
493
−52.240
−34.603
−7.268
1.00
40.76
C


ATOM
15801
O
PHE
B
493
−52.686
−35.733
−7.419
1.00
40.35
O


ATOM
15803
N
ALA
B
494
−52.943
−33.584
−6.790
1.00
40.72
N


ATOM
15804
CA
ALA
B
494
−54.355
−33.658
−6.471
1.00
40.68
C


ATOM
15806
CB
ALA
B
494
−54.786
−32.362
−5.770
1.00
40.70
C


ATOM
15810
C
ALA
B
494
−55.175
−33.851
−7.744
1.00
40.59
C


ATOM
15811
O
ALA
B
494
−55.013
−33.099
−8.716
1.00
40.95
O


ATOM
15813
N
LYS
B
495
−56.071
−34.835
−7.733
1.00
40.12
N


ATOM
15814
CA
LYS
B
495
−57.000
−35.047
−8.854
1.00
39.67
C


ATOM
15816
CB
LYS
B
495
−58.008
−36.166
−8.520
1.00
39.98
C


ATOM
15819
CG
LYS
B
495
−57.388
−37.590
−8.496
1.00
40.57
C


ATOM
15822
CD
LYS
B
495
−58.468
−38.688
−8.419
1.00
41.18
C


ATOM
15825
CE
LYS
B
495
−57.878
−40.065
−8.132
1.00
41.28
C


ATOM
15828
NZ
LYS
B
495
−58.736
−41.159
−8.671
1.00
41.39
N


ATOM
15832
C
LYS
B
495
−57.717
−33.752
−9.333
1.00
38.82
C


ATOM
15833
O
LYS
B
495
−57.847
−33.535
−10.537
1.00
38.70
O


ATOM
15835
N
PRO
B
496
−58.169
−32.890
−8.397
1.00
37.71
N


ATOM
15836
CA
PRO
B
496
−58.701
−31.551
−8.687
1.00
36.80
C


ATOM
15838
CB
PRO
B
496
−58.820
−30.937
−7.304
1.00
36.94
C


ATOM
15841
CG
PRO
B
496
−59.255
−32.073
−6.483
1.00
37.90
C


ATOM
15844
CD
PRO
B
496
−58.534
−33.294
−7.030
1.00
37.86
C


ATOM
15847
C
PRO
B
496
−57.845
−30.625
−9.536
1.00
35.57
C


ATOM
15848
O
PRO
B
496
−58.389
−29.693
−10.139
1.00
35.93
O


ATOM
15849
N
PHE
B
497
−56.527
−30.836
−9.546
1.00
33.52
N


ATOM
15850
CA
PHE
B
497
−55.654
−30.083
−10.439
1.00
31.34
C


ATOM
15852
CB
PHE
B
497
−54.355
−29.710
−9.750
1.00
30.86
C


ATOM
15855
CG
PHE
B
497
−53.394
−28.989
−10.634
1.00
29.04
C


ATOM
15856
CD1
PHE
B
497
−53.639
−27.700
−11.022
1.00
27.89
C


ATOM
15858
CE1
PHE
B
497
−52.752
−27.028
−11.834
1.00
27.39
C


ATOM
15860
CZ
PHE
B
497
−51.599
−27.653
−12.259
1.00
27.40
C


ATOM
15862
CE2
PHE
B
497
−51.341
−28.941
−11.877
1.00
27.44
C


ATOM
15864
CD2
PHE
B
497
−52.236
−29.604
−11.074
1.00
28.18
C


ATOM
15866
C
PHE
B
497
−55.390
−30.870
−11.714
1.00
30.24
C


ATOM
15867
O
PHE
B
497
−55.301
−30.266
−12.786
1.00
30.39
O


ATOM
15869
N
VAL
B
498
−55.295
−32.200
−11.630
1.00
28.50
N


ATOM
15870
CA
VAL
B
498
−55.076
−32.983
−12.854
1.00
27.54
C


ATOM
15872
CB
VAL
B
498
−54.778
−34.472
−12.611
1.00
27.34
C


ATOM
15874
CG1
VAL
B
498
−56.027
−35.194
−12.262
1.00
28.14
C


ATOM
15878
CG2
VAL
B
498
−53.731
−34.649
−11.530
1.00
27.04
C


ATOM
15882
C
VAL
B
498
−56.277
−32.857
−13.784
1.00
26.60
C


ATOM
15883
O
VAL
B
498
−56.129
−32.864
−14.995
1.00
26.33
O


ATOM
15885
N
GLU
B
499
−57.468
−32.731
−13.214
1.00
25.87
N


ATOM
15886
CA
GLU
B
499
−58.662
−32.484
−14.022
1.00
25.20
C


ATOM
15888
CB
GLU
B
499
−59.975
−32.639
−13.206
1.00
25.26
C


ATOM
15891
CG
GLU
B
499
−61.287
−32.694
−14.028
1.00
25.16
C


ATOM
15894
CD
GLU
B
499
−61.422
−33.922
−14.973
1.00
26.79
C


ATOM
15895
OE1
GLU
B
499
−60.607
−34.880
−14.928
1.00
26.39
O


ATOM
15896
OE2
GLU
B
499
−62.379
−33.926
−15.785
1.00
27.83
O


ATOM
15897
C
GLU
B
499
−58.542
−31.097
−14.641
1.00
24.27
C


ATOM
15898
O
GLU
B
499
−58.819
−30.948
−15.832
1.00
24.32
O


ATOM
15900
N
THR
B
500
−58.098
−30.093
−13.882
1.00
22.90
N


ATOM
15901
CA
THR
B
500
−58.027
−28.764
−14.487
1.00
22.29
C


ATOM
15903
CB
THR
B
500
−57.908
−27.592
−13.488
1.00
22.43
C


ATOM
15905
OG1
THR
B
500
−56.536
−27.259
−13.274
1.00
22.65
O


ATOM
15907
CG2
THR
B
500
−58.614
−27.899
−12.183
1.00
22.84
C


ATOM
15911
C
THR
B
500
−56.940
−28.671
−15.565
1.00
21.20
C


ATOM
15912
O
THR
B
500
−57.120
−27.939
−16.536
1.00
21.06
O


ATOM
15914
N
ALA
B
501
−55.845
−29.414
−15.422
1.00
19.88
N


ATOM
15915
CA
ALA
B
501
−54.906
−29.574
−16.537
1.00
19.24
C


ATOM
15917
CB
ALA
B
501
−53.704
−30.393
−16.115
1.00
19.22
C


ATOM
15921
C
ALA
B
501
−55.581
−30.226
−17.757
1.00
18.68
C


ATOM
15922
O
ALA
B
501
−55.427
−29.768
−18.881
1.00
18.32
O


ATOM
15924
N
ILE
B
502
−56.324
−31.300
−17.523
1.00
18.11
N


ATOM
15925
CA
ILE
B
502
−57.023
−31.988
−18.591
1.00
17.68
C


ATOM
15927
CB
ILE
B
502
−57.750
−33.251
−18.059
1.00
17.63
C


ATOM
15929
CG1
ILE
B
502
−56.713
−34.350
−17.769
1.00
18.16
C


ATOM
15932
CD1
ILE
B
502
−57.253
−35.645
−17.115
1.00
17.02
C


ATOM
15936
CG2
ILE
B
502
−58.756
−33.762
−19.063
1.00
16.93
C


ATOM
15940
C
ILE
B
502
−57.990
−31.030
−19.289
1.00
17.50
C


ATOM
15941
O
ILE
B
502
−58.153
−31.071
−20.512
1.00
17.62
O


ATOM
15943
N
ASN
B
503
−58.604
−30.139
−18.524
1.00
17.23
N


ATOM
15944
CA
ASN
B
503
−59.545
−29.184
−19.102
1.00
17.07
C


ATOM
15946
CB
ASN
B
503
−60.184
−28.327
−18.016
1.00
17.21
C


ATOM
15949
CG
ASN
B
503
−61.155
−29.094
−17.176
1.00
17.29
C


ATOM
15950
OD1
ASN
B
503
−61.721
−30.097
−17.609
1.00
16.66
O


ATOM
15951
ND2
ASN
B
503
−61.365
−28.621
−15.958
1.00
19.16
N


ATOM
15954
C
ASN
B
503
−58.920
−28.270
−20.149
1.00
16.59
C


ATOM
15955
O
ASN
B
503
−59.611
−27.811
−21.071
1.00
16.67
O


ATOM
15957
N
LEU
B
504
−57.629
−27.992
−20.014
1.00
15.79
N


ATOM
15958
CA
LEU
B
504
−56.948
−27.204
−21.031
1.00
15.44
C


ATOM
15960
CB
LEU
B
504
−55.530
−26.865
−20.609
1.00
15.07
C


ATOM
15963
CG
LEU
B
504
−54.933
−25.837
−21.552
1.00
14.21
C


ATOM
15965
CD1
LEU
B
504
−54.246
−24.726
−20.800
1.00
14.21
C


ATOM
15969
CD2
LEU
B
504
−54.000
−26.538
−22.462
1.00
14.27
C


ATOM
15973
C
LEU
B
504
−56.954
−27.933
−22.375
1.00
15.59
C


ATOM
15974
O
LEU
B
504
−57.177
−27.323
−23.414
1.00
15.30
O


ATOM
15976
N
ALA
B
505
−56.732
−29.242
−22.341
1.00
15.89
N


ATOM
15977
CA
ALA
B
505
−56.914
−30.073
−23.514
1.00
16.23
C


ATOM
15979
CB
ALA
B
505
−56.608
−31.483
−23.188
1.00
16.05
C


ATOM
15983
C
ALA
B
505
−58.352
−29.955
−24.010
1.00
16.69
C


ATOM
15984
O
ALA
B
505
−58.598
−29.738
−25.204
1.00
16.88
O


ATOM
15986
N
ARG
B
506
−59.300
−30.071
−23.090
1.00
16.88
N


ATOM
15987
CA
ARG
B
506
−60.709
−29.972
−23.457
1.00
17.20
C


ATOM
15989
CB
ARG
B
506
−61.630
−30.170
−22.248
1.00
17.34
C


ATOM
15992
CG
ARG
B
506
−61.549
−31.537
−21.614
1.00
17.46
C


ATOM
15995
CD
ARG
B
506
−62.837
−31.917
−20.962
1.00
17.48
C


ATOM
15998
NE
ARG
B
506
−62.783
−33.280
−20.435
1.00
18.37
N


ATOM
16000
CZ
ARG
B
506
−62.404
−33.613
−19.201
1.00
19.03
C


ATOM
16001
NH1
ARG
B
506
−62.008
−32.690
−18.324
1.00
18.83
N


ATOM
16004
NH2
ARG
B
506
−62.415
−34.892
−18.841
1.00
19.72
N


ATOM
16007
C
ARG
B
506
−61.038
−28.646
−24.114
1.00
17.32
C


ATOM
16008
O
ARG
B
506
−61.754
−28.622
−25.111
1.00
17.41
O


ATOM
16010
N
GLN
B
507
−60.540
−27.543
−23.564
1.00
17.59
N


ATOM
16011
CA
GLN
B
507
−60.864
−26.233
−24.139
1.00
18.05
C


ATOM
16013
CB
GLN
B
507
−60.382
−25.090
−23.258
1.00
18.16
C


ATOM
16016
CG
GLN
B
507
−60.798
−23.709
−23.764
1.00
17.57
C


ATOM
16019
CD
GLN
B
507
−62.291
−23.508
−23.723
1.00
17.42
C


ATOM
16020
OE1
GLN
B
507
−62.953
−23.959
−22.800
1.00
17.55
O


ATOM
16021
NE2
GLN
B
507
−62.832
−22.823
−24.722
1.00
17.56
N


ATOM
16024
C
GLN
B
507
−60.258
−26.060
−25.521
1.00
18.49
C


ATOM
16025
O
GLN
B
507
−60.855
−25.413
−26.380
1.00
18.73
O


ATOM
16027
N
SER
B
508
−59.066
−26.619
−25.724
1.00
18.85
N


ATOM
16028
CA
SER
B
508
−58.427
−26.621
−27.037
1.00
19.07
C


ATOM
16030
CB
SER
B
508
−57.108
−27.364
−26.969
1.00
19.04
C


ATOM
16033
OG
SER
B
508
−56.304
−26.772
−25.979
1.00
20.10
O


ATOM
16035
C
SER
B
508
−59.305
−27.296
−28.065
1.00
19.12
C


ATOM
16036
O
SER
B
508
−59.438
−26.827
−29.187
1.00
19.00
O


ATOM
16038
N
HIS
B
509
−59.905
−28.409
−27.674
1.00
19.26
N


ATOM
16039
CA
HIS
B
509
−60.814
−29.095
−28.559
1.00
19.33
C


ATOM
16041
CB
HIS
B
509
−61.275
−30.412
−27.959
1.00
19.30
C


ATOM
16044
CG
HIS
B
509
−60.263
−31.501
−28.067
1.00
18.71
C


ATOM
16045
ND1
HIS
B
509
−59.941
−32.089
−29.267
1.00
18.57
N


ATOM
16047
CE1
HIS
B
509
−59.020
−33.012
−29.064
1.00
19.07
C


ATOM
16049
NE2
HIS
B
509
−58.738
−33.044
−27.774
1.00
18.52
N


ATOM
16051
CD2
HIS
B
509
−59.499
−32.105
−27.129
1.00
18.43
C


ATOM
16053
C
HIS
B
509
−62.017
−28.251
−28.878
1.00
19.60
C


ATOM
16054
O
HIS
B
509
−62.491
−28.286
−29.969
1.00
19.68
O


ATOM
16056
N
CYS
B
510
−62.540
−27.502
−27.932
1.00
20.12
N


ATOM
16057
CA
CYS
B
510
−63.748
−26.738
−28.216
1.00
20.63
C


ATOM
16059
CB
CYS
B
510
−64.493
−26.492
−26.915
1.00
20.70
C


ATOM
16062
SG
CYS
B
510
−64.856
−28.020
−26.065
1.00
22.93
S


ATOM
16064
C
CYS
B
510
−63.465
−25.422
−28.950
1.00
20.55
C


ATOM
16065
O
CYS
B
510
−64.338
−24.881
−29.622
1.00
20.16
O


ATOM
16067
N
THR
B
511
−62.236
−24.933
−28.827
1.00
20.89
N


ATOM
16068
CA
THR
B
511
−61.833
−23.654
−29.378
1.00
21.32
C


ATOM
16070
CB
THR
B
511
−60.682
−23.107
−28.550
1.00
20.76
C


ATOM
16072
OG1
THR
B
511
−61.208
−22.619
−27.324
1.00
19.50
O


ATOM
16074
CG2
THR
B
511
−59.961
−21.994
−29.253
1.00
19.79
C


ATOM
16078
C
THR
B
511
−61.417
−23.735
−30.851
1.00
23.11
C


ATOM
16079
O
THR
B
511
−61.910
−22.992
−31.694
1.00
22.75
O


ATOM
16081
N
TYR
B
512
−60.501
−24.646
−31.152
1.00
25.46
N


ATOM
16082
CA
TYR
B
512
−59.867
−24.704
−32.462
1.00
27.09
C


ATOM
16084
CB
TYR
B
512
−58.397
−25.107
−32.327
1.00
27.16
C


ATOM
16087
CG
TYR
B
512
−57.598
−24.056
−31.582
1.00
27.26
C


ATOM
16088
CD1
TYR
B
512
−57.300
−22.834
−32.182
1.00
28.18
C


ATOM
16090
CE1
TYR
B
512
−56.583
−21.844
−31.511
1.00
27.98
C


ATOM
16092
CZ
TYR
B
512
−56.164
−22.068
−30.226
1.00
27.72
C


ATOM
16093
OH
TYR
B
512
−55.462
−21.072
−29.584
1.00
27.09
O


ATOM
16095
CE2
TYR
B
512
−56.453
−23.281
−29.602
1.00
27.52
C


ATOM
16097
CD2
TYR
B
512
−57.171
−24.262
−30.278
1.00
26.68
C


ATOM
16099
C
TYR
B
512
−60.633
−25.623
−33.382
1.00
28.79
C


ATOM
16100
O
TYR
B
512
−61.050
−25.179
−34.444
1.00
28.98
O


ATOM
16102
N
HIS
B
513
−60.789
−26.893
−32.991
1.00
31.06
N


ATOM
16103
CA
HIS
B
513
−61.865
−27.800
−33.492
1.00
33.07
C


ATOM
16105
CB
HIS
B
513
−62.740
−28.208
−32.265
1.00
33.66
C


ATOM
16108
CG
HIS
B
513
−64.181
−28.602
−32.535
1.00
34.73
C


ATOM
16109
ND1
HIS
B
513
−65.035
−27.903
−33.367
1.00
35.70
N


ATOM
16111
CE1
HIS
B
513
−66.234
−28.464
−33.343
1.00
35.40
C


ATOM
16113
NE2
HIS
B
513
−66.206
−29.469
−32.491
1.00
35.10
N


ATOM
16115
CD2
HIS
B
513
−64.943
−29.565
−31.956
1.00
35.07
C


ATOM
16117
C
HIS
B
513
−62.692
−27.179
−34.623
1.00
34.03
C


ATOM
16118
O
HIS
B
513
−63.003
−27.848
−35.627
1.00
34.55
O


ATOM
16120
N
ASN
B
514
−63.047
−25.906
−34.444
1.00
34.65
N


ATOM
16121
CA
ASN
B
514
−63.713
−25.123
−35.483
1.00
35.15
C


ATOM
16123
CB
ASN
B
514
−63.845
−23.643
−35.070
1.00
35.01
C


ATOM
16126
CG
ASN
B
514
−64.704
−23.456
−33.820
1.00
33.48
C


ATOM
16127
OD1
ASN
B
514
−64.787
−24.343
−32.968
1.00
30.92
O


ATOM
16128
ND2
ASN
B
514
−65.327
−22.295
−33.703
1.00
32.16
N


ATOM
16131
C
ASN
B
514
−63.192
−25.274
−36.942
1.00
36.08
C


ATOM
16132
O
ASN
B
514
−62.010
−25.051
−37.254
1.00
35.69
O


ATOM
16134
N
GLY
B
515
−64.119
−25.765
−37.772
1.00
37.15
N


ATOM
16135
CA
GLY
B
515
−64.224
−25.493
−39.177
1.00
37.94
C


ATOM
16138
C
GLY
B
515
−65.269
−24.384
−39.284
1.00
39.01
C


ATOM
16139
O
GLY
B
515
−64.904
−23.286
−39.702
1.00
39.58
O


ATOM
16141
N
ASP
B
516
−66.550
−24.585
−38.909
1.00
39.95
N


ATOM
16142
CA
ASP
B
516
−67.167
−25.810
−38.349
1.00
40.65
C


ATOM
16144
CB
ASP
B
516
−67.296
−25.652
−36.836
1.00
41.09
C


ATOM
16147
CG
ASP
B
516
−66.318
−26.511
−36.063
1.00
43.63
C


ATOM
16148
OD1
ASP
B
516
−65.697
−27.434
−36.662
1.00
45.83
O


ATOM
16149
OD2
ASP
B
516
−66.152
−26.246
−34.843
1.00
47.12
O


ATOM
16150
C
ASP
B
516
−68.603
−26.067
−38.853
1.00
40.78
C


ATOM
16151
O
ASP
B
516
−69.134
−25.301
−39.660
1.00
41.28
O


ATOM
16153
N
ALA
B
517
−69.222
−27.141
−38.349
1.00
40.66
N


ATOM
16154
CA
ALA
B
517
−70.675
−27.396
−38.468
1.00
40.60
C


ATOM
16156
CB
ALA
B
517
−71.422
−26.622
−37.359
1.00
40.27
C


ATOM
16160
C
ALA
B
517
−71.295
−27.106
−39.862
1.00
40.79
C


ATOM
16161
O
ALA
B
517
−70.654
−27.296
−40.899
1.00
40.67
O


ATOM
16163
N
HIS
B
518
−72.562
−26.696
−39.874
1.00
41.13
N


ATOM
16164
CA
HIS
B
518
−73.162
−26.033
−41.039
1.00
41.60
C


ATOM
16166
CB
HIS
B
518
−74.446
−26.756
−41.480
1.00
42.24
C


ATOM
16169
CG
HIS
B
518
−74.207
−28.179
−41.910
1.00
45.54
C


ATOM
16170
ND1
HIS
B
518
−73.441
−28.509
−43.012
1.00
48.51
N


ATOM
16172
CE1
HIS
B
518
−73.392
−29.825
−43.140
1.00
48.79
C


ATOM
16174
NE2
HIS
B
518
−74.092
−30.364
−42.156
1.00
49.12
N


ATOM
16176
CD2
HIS
B
518
−74.608
−29.357
−41.369
1.00
48.28
C


ATOM
16178
C
HIS
B
518
−73.377
−24.545
−40.686
1.00
40.84
C


ATOM
16179
O
HIS
B
518
−74.487
−23.997
−40.769
1.00
40.60
O


ATOM
16181
N
THR
B
519
−72.261
−23.932
−40.275
1.00
40.10
N


ATOM
16182
CA
THR
B
519
−72.143
−22.521
−39.878
1.00
39.39
C


ATOM
16184
CB
THR
B
519
−72.561
−22.263
−38.393
1.00
39.40
C


ATOM
16186
OG1
THR
B
519
−71.823
−23.131
−37.517
1.00
39.55
O


ATOM
16188
CG2
THR
B
519
−74.074
−22.460
−38.178
1.00
38.98
C


ATOM
16192
C
THR
B
519
−70.654
−22.164
−40.059
1.00
38.81
C


ATOM
16193
O
THR
B
519
−69.800
−23.050
−40.067
1.00
38.48
O


ATOM
16195
N
SER
B
520
−70.338
−20.881
−40.199
1.00
38.19
N


ATOM
16196
CA
SER
B
520
−68.959
−20.453
−40.521
1.00
37.77
C


ATOM
16198
CB
SER
B
520
−68.983
−18.976
−40.960
1.00
37.76
C


ATOM
16201
OG
SER
B
520
−68.760
−18.106
−39.870
1.00
38.49
O


ATOM
16203
C
SER
B
520
−67.974
−20.724
−39.340
1.00
37.24
C


ATOM
16204
O
SER
B
520
−68.394
−21.267
−38.320
1.00
36.89
O


ATOM
16206
N
PRO
B
521
−66.671
−20.350
−39.472
1.00
36.96
N


ATOM
16207
CA
PRO
B
521
−65.692
−20.638
−38.396
1.00
36.80
C


ATOM
16209
CB
PRO
B
521
−64.320
−20.354
−39.041
1.00
36.68
C


ATOM
16212
CG
PRO
B
521
−64.599
−19.593
−40.309
1.00
37.19
C


ATOM
16215
CD
PRO
B
521
−66.091
−19.469
−40.507
1.00
37.10
C


ATOM
16218
C
PRO
B
521
−65.915
−19.763
−37.168
1.00
36.68
C


ATOM
16219
O
PRO
B
521
−66.233
−20.289
−36.105
1.00
36.90
O


ATOM
16220
N
ASP
B
522
−65.744
−18.445
−37.298
1.00
36.61
N


ATOM
16221
CA
ASP
B
522
−66.373
−17.528
−36.347
1.00
36.58
C


ATOM
16223
CB
ASP
B
522
−65.938
−16.070
−36.543
1.00
36.82
C


ATOM
16226
CG
ASP
B
522
−64.456
−15.846
−36.243
1.00
37.94
C


ATOM
16227
OD1
ASP
B
522
−63.848
−16.627
−35.473
1.00
39.62
O


ATOM
16228
OD2
ASP
B
522
−63.891
−14.876
−36.790
1.00
39.40
O


ATOM
16229
C
ASP
B
522
−67.836
−17.718
−36.681
1.00
36.12
C


ATOM
16230
O
ASP
B
522
−68.150
−18.306
−37.702
1.00
35.98
O


ATOM
16232
N
GLU
B
523
−68.732
−17.249
−35.830
1.00
35.69
N


ATOM
16233
CA
GLU
B
523
−70.152
−17.605
−35.927
1.00
35.46
C


ATOM
16235
CB
GLU
B
523
−70.703
−17.560
−37.379
1.00
35.51
C


ATOM
16238
CG
GLU
B
523
−70.535
−16.175
−38.061
1.00
36.11
C


ATOM
16241
CD
GLU
B
523
−70.854
−16.136
−39.572
1.00
36.72
C


ATOM
16242
OE1
GLU
B
523
−71.848
−16.767
−40.013
1.00
37.20
O


ATOM
16243
OE2
GLU
B
523
−70.101
−15.452
−40.314
1.00
35.53
O


ATOM
16244
C
GLU
B
523
−70.440
−18.943
−35.216
1.00
34.97
C


ATOM
16245
O
GLU
B
523
−71.557
−19.148
−34.765
1.00
35.02
O


ATOM
16247
N
LEU
B
524
−69.462
−19.846
−35.099
1.00
34.59
N


ATOM
16248
CA
LEU
B
524
−69.503
−20.832
−34.004
1.00
34.45
C


ATOM
16250
CB
LEU
B
524
−68.735
−22.136
−34.286
1.00
34.21
C


ATOM
16253
CG
LEU
B
524
−69.530
−23.434
−34.508
1.00
33.92
C


ATOM
16255
CD1
LEU
B
524
−68.657
−24.627
−34.227
1.00
32.21
C


ATOM
16259
CD2
LEU
B
524
−70.794
−23.520
−33.641
1.00
34.06
C


ATOM
16263
C
LEU
B
524
−68.901
−20.162
−32.791
1.00
34.38
C


ATOM
16264
O
LEU
B
524
−69.535
−20.067
−31.741
1.00
34.49
O


ATOM
16266
N
THR
B
525
−67.670
−19.688
−32.958
1.00
34.30
N


ATOM
16267
CA
THR
B
525
−66.913
−19.073
−31.874
1.00
34.23
C


ATOM
16269
CB
THR
B
525
−65.570
−18.524
−32.380
1.00
34.04
C


ATOM
16271
OG1
THR
B
525
−64.894
−19.549
−33.112
1.00
33.83
O


ATOM
16273
CG2
THR
B
525
−64.689
−18.087
−31.227
1.00
33.37
C


ATOM
16277
C
THR
B
525
−67.702
−17.960
−31.184
1.00
34.46
C


ATOM
16278
O
THR
B
525
−67.684
−17.858
−29.955
1.00
34.57
O


ATOM
16280
N
ARG
B
526
−68.401
−17.144
−31.969
1.00
34.54
N


ATOM
16281
CA
ARG
B
526
−69.200
−16.056
−31.418
1.00
34.60
C


ATOM
16283
CB
ARG
B
526
−69.571
−15.059
−32.513
1.00
35.04
C


ATOM
16286
CG
ARG
B
526
−70.256
−13.800
−32.013
1.00
37.13
C


ATOM
16289
CD
ARG
B
526
−70.143
−12.656
−33.026
1.00
39.87
C


ATOM
16292
NE
ARG
B
526
−70.353
−13.075
−34.420
1.00
42.28
N


ATOM
16294
CZ
ARG
B
526
−71.543
−13.292
−35.000
1.00
44.26
C


ATOM
16295
NH1
ARG
B
526
−72.685
−13.159
−34.320
1.00
44.66
N


ATOM
16298
NH2
ARG
B
526
−71.593
−13.658
−36.279
1.00
44.49
N


ATOM
16301
C
ARG
B
526
−70.448
−16.593
−30.735
1.00
33.91
C


ATOM
16302
O
ARG
B
526
−70.848
−16.074
−29.704
1.00
33.74
O


ATOM
16304
N
LYS
B
527
−71.048
−17.637
−31.306
1.00
33.39
N


ATOM
16305
CA
LYS
B
527
−72.240
−18.252
−30.722
1.00
33.15
C


ATOM
16307
CB
LYS
B
527
−72.837
−19.332
−31.639
1.00
33.42
C


ATOM
16310
CG
LYS
B
527
−73.898
−18.804
−32.609
1.00
34.55
C


ATOM
16313
CD
LYS
B
527
−74.643
−19.919
−33.358
1.00
35.43
C


ATOM
16316
CE
LYS
B
527
−75.230
−19.383
−34.667
1.00
36.11
C


ATOM
16319
NZ
LYS
B
527
−76.076
−20.372
−35.384
1.00
37.04
N


ATOM
16323
C
LYS
B
527
−71.925
−18.865
−29.378
1.00
32.36
C


ATOM
16324
O
LYS
B
527
−72.589
−18.584
−28.387
1.00
32.56
O


ATOM
16326
N
ARG
B
528
−70.909
−19.713
−29.358
1.00
31.53
N


ATOM
16327
CA
ARG
B
528
−70.502
−20.401
−28.138
1.00
30.77
C


ATOM
16329
CB
ARG
B
528
−69.283
−21.286
−28.414
1.00
30.55
C


ATOM
16332
CG
ARG
B
528
−69.624
−22.519
−29.252
1.00
29.48
C


ATOM
16335
CD
ARG
B
528
−68.418
−23.417
−29.493
1.00
28.05
C


ATOM
16338
NE
ARG
B
528
−68.811
−24.774
−29.880
1.00
26.55
N


ATOM
16340
CZ
ARG
B
528
−67.968
−25.719
−30.288
1.00
25.87
C


ATOM
16341
NH1
ARG
B
528
−66.668
−25.472
−30.385
1.00
26.33
N


ATOM
16344
NH2
ARG
B
528
−68.424
−26.920
−30.615
1.00
25.49
N


ATOM
16347
C
ARG
B
528
−70.225
−19.415
−27.004
1.00
30.42
C


ATOM
16348
O
ARG
B
528
−70.721
−19.582
−25.885
1.00
30.30
O


ATOM
16350
N
VAL
B
529
−69.455
−18.376
−27.307
1.00
29.96
N


ATOM
16351
CA
VAL
B
529
−69.194
−17.312
−26.342
1.00
29.47
C


ATOM
16353
CB
VAL
B
529
−68.261
−16.227
−26.932
1.00
29.40
C


ATOM
16355
CG1
VAL
B
529
−68.269
−14.968
−26.081
1.00
29.18
C


ATOM
16359
CG2
VAL
B
529
−66.845
−16.782
−27.062
1.00
28.90
C


ATOM
16363
C
VAL
B
529
−70.516
−16.723
−25.841
1.00
28.98
C


ATOM
16364
O
VAL
B
529
−70.759
−16.682
−24.641
1.00
28.97
O


ATOM
16366
N
LEU
B
530
−71.384
−16.308
−26.752
1.00
28.54
N


ATOM
16367
CA
LEU
B
530
−72.709
−15.824
−26.353
1.00
28.23
C


ATOM
16369
CB
LEU
B
530
−73.631
−15.604
−27.559
1.00
28.09
C


ATOM
16372
CG
LEU
B
530
−73.767
−14.150
−28.007
1.00
28.15
C


ATOM
16374
CD1
LEU
B
530
−72.412
−13.570
−28.406
1.00
27.99
C


ATOM
16378
CD2
LEU
B
530
−74.784
−14.034
−29.150
1.00
28.85
C


ATOM
16382
C
LEU
B
530
−73.389
−16.767
−25.367
1.00
27.88
C


ATOM
16383
O
LEU
B
530
−73.932
−16.319
−24.363
1.00
28.11
O


ATOM
16385
N
SER
B
531
−73.352
−18.066
−25.652
1.00
27.28
N


ATOM
16386
CA
SER
B
531
−74.081
−19.049
−24.849
1.00
26.68
C


ATOM
16388
CB
SER
B
531
−74.209
−20.363
−25.612
1.00
26.68
C


ATOM
16391
OG
SER
B
531
−72.970
−21.033
−25.681
1.00
26.67
O


ATOM
16393
C
SER
B
531
−73.410
−19.312
−23.514
1.00
26.20
C


ATOM
16394
O
SER
B
531
−74.076
−19.550
−22.511
1.00
25.86
O


ATOM
16396
N
VAL
B
532
−72.085
−19.281
−23.512
1.00
25.86
N


ATOM
16397
CA
VAL
B
532
−71.320
−19.562
−22.306
1.00
25.51
C


ATOM
16399
CB
VAL
B
532
−69.891
−20.055
−22.651
1.00
25.34
C


ATOM
16401
CG1
VAL
B
532
−68.955
−19.932
−21.460
1.00
24.01
C


ATOM
16405
CG2
VAL
B
532
−69.950
−21.484
−23.158
1.00
24.95
C


ATOM
16409
C
VAL
B
532
−71.271
−18.357
−21.373
1.00
25.52
C


ATOM
16410
O
VAL
B
532
−71.377
−18.523
−20.164
1.00
25.82
O


ATOM
16412
N
ILE
B
533
−71.137
−17.158
−21.935
1.00
25.39
N


ATOM
16413
CA
ILE
B
533
−70.875
−15.954
−21.151
1.00
25.48
C


ATOM
16415
CB
ILE
B
533
−69.666
−15.185
−21.732
1.00
25.30
C


ATOM
16417
CG1
ILE
B
533
−68.375
−15.928
−21.451
1.00
24.91
C


ATOM
16420
CD1
ILE
B
533
−68.118
−16.120
−19.989
1.00
25.00
C


ATOM
16424
CG2
ILE
B
533
−69.553
−13.797
−21.130
1.00
25.81
C


ATOM
16428
C
ILE
B
533
−72.065
−14.983
−21.030
1.00
25.77
C


ATOM
16429
O
ILE
B
533
−72.537
−14.707
−19.928
1.00
25.77
O


ATOM
16431
N
THR
B
534
−72.541
−14.442
−22.142
1.00
25.98
N


ATOM
16432
CA
THR
B
534
−73.446
−13.295
−22.064
1.00
26.31
C


ATOM
16434
CB
THR
B
534
−73.094
−12.253
−23.143
1.00
26.21
C


ATOM
16436
OG1
THR
B
534
−72.914
−12.909
−24.397
1.00
26.72
O


ATOM
16438
CG2
THR
B
534
−71.795
−11.535
−22.781
1.00
25.97
C


ATOM
16442
C
THR
B
534
−74.961
−13.621
−22.064
1.00
26.50
C


ATOM
16443
O
THR
B
534
−75.713
−12.974
−21.344
1.00
26.67
O


ATOM
16445
N
GLU
B
535
−75.410
−14.616
−22.829
1.00
26.55
N


ATOM
16446
CA
GLU
B
535
−76.851
−14.892
−22.961
1.00
26.50
C


ATOM
16448
CB
GLU
B
535
−77.197
−15.197
−24.426
1.00
26.68
C


ATOM
16451
CG
GLU
B
535
−77.226
−13.936
−25.296
1.00
27.68
C


ATOM
16454
CD
GLU
B
535
−78.193
−14.014
−26.475
1.00
28.75
C


ATOM
16455
OE1
GLU
B
535
−79.370
−13.588
−26.330
1.00
27.72
O


ATOM
16456
OE2
GLU
B
535
−77.762
−14.492
−27.549
1.00
30.06
O


ATOM
16457
C
GLU
B
535
−77.355
−16.019
−22.056
1.00
26.19
C


ATOM
16458
O
GLU
B
535
−77.021
−17.171
−22.279
1.00
26.15
O


ATOM
16460
N
PRO
B
536
−78.186
−15.697
−21.047
1.00
26.05
N


ATOM
16461
CA
PRO
B
536
−78.692
−16.763
−20.196
1.00
26.05
C


ATOM
16463
CB
PRO
B
536
−79.495
−16.018
−19.118
1.00
25.87
C


ATOM
16466
CG
PRO
B
536
−79.140
−14.621
−19.239
1.00
25.88
C


ATOM
16469
CD
PRO
B
536
−78.760
−14.402
−20.660
1.00
26.21
C


ATOM
16472
C
PRO
B
536
−79.608
−17.717
−20.943
1.00
26.14
C


ATOM
16473
O
PRO
B
536
−80.173
−17.359
−21.973
1.00
25.99
O


ATOM
16474
N
ILE
B
537
−79.746
−18.924
−20.411
1.00
26.38
N


ATOM
16475
CA
ILE
B
537
−80.662
−19.905
−20.960
1.00
26.58
C


ATOM
16477
CB
ILE
B
537
−80.443
−21.292
−20.333
1.00
26.47
C


ATOM
16479
CG1
ILE
B
537
−79.023
−21.789
−20.600
1.00
26.38
C


ATOM
16482
CD1
ILE
B
537
−78.709
−23.102
−19.907
1.00
26.40
C


ATOM
16486
CG2
ILE
B
537
−81.430
−22.300
−20.890
1.00
26.71
C


ATOM
16490
C
ILE
B
537
−82.072
−19.420
−20.657
1.00
26.93
C


ATOM
16491
O
ILE
B
537
−82.347
−18.965
−19.545
1.00
26.81
O


ATOM
16493
N
LEU
B
538
−82.963
−19.491
−21.641
1.00
27.36
N


ATOM
16494
CA
LEU
B
538
−84.309
−18.984
−21.439
1.00
27.68
C


ATOM
16496
CB
LEU
B
538
−85.181
−19.094
−22.698
1.00
27.75
C


ATOM
16499
CG
LEU
B
538
−84.782
−18.291
−23.949
1.00
27.47
C


ATOM
16501
CD1
LEU
B
538
−85.992
−18.109
−24.844
1.00
27.11
C


ATOM
16505
CD2
LEU
B
538
−84.164
−16.933
−23.621
1.00
26.99
C


ATOM
16509
C
LEU
B
538
−84.919
−19.752
−20.288
1.00
28.01
C


ATOM
16512
N
PRO
B
539
−85.645
−19.046
−19.421
1.00
28.69
N


ATOM
16513
CA
PRO
B
539
−86.028
−19.631
−18.152
1.00
29.05
C


ATOM
16515
CB
PRO
B
539
−86.640
−18.452
−17.397
1.00
29.08
C


ATOM
16518
CG
PRO
B
539
−87.144
−17.548
−18.447
1.00
28.86
C


ATOM
16521
CD
PRO
B
539
−86.298
−17.748
−19.660
1.00
28.62
C


ATOM
16524
C
PRO
B
539
−87.035
−20.767
−18.259
1.00
29.51
C


ATOM
16525
O
PRO
B
539
−87.665
−20.970
−19.297
1.00
29.23
O


ATOM
16526
N
PHE
B
540
−87.164
−21.500
−17.159
1.00
30.25
N


ATOM
16527
CA
PHE
B
540
−88.089
−22.613
−17.070
1.00
30.56
C


ATOM
16529
CB
PHE
B
540
−87.971
−23.297
−15.708
1.00
30.73
C


ATOM
16532
CG
PHE
B
540
−88.848
−24.499
−15.567
1.00
30.49
C


ATOM
16533
CD1
PHE
B
540
−89.910
−24.505
−14.683
1.00
30.32
C


ATOM
16535
CE1
PHE
B
540
−90.718
−25.614
−14.568
1.00
30.59
C


ATOM
16537
CZ
PHE
B
540
−90.475
−26.722
−15.346
1.00
30.55
C


ATOM
16539
CE2
PHE
B
540
−89.420
−26.720
−16.234
1.00
30.68
C


ATOM
16541
CD2
PHE
B
540
−88.619
−25.617
−16.342
1.00
30.43
C


ATOM
16543
C
PHE
B
540
−89.507
−22.120
−17.257
1.00
30.76
C


ATOM
16544
O
PHE
B
540
−89.967
−21.257
−16.508
1.00
30.59
O


ATOM
16546
N
GLU
B
541
−90.184
−22.677
−18.259
1.00
31.13
N


ATOM
16547
CA
GLU
B
541
−91.553
−22.289
−18.608
1.00
31.45
C


ATOM
16549
CB
GLU
B
541
−91.525
−21.186
−19.680
1.00
31.60
C


ATOM
16552
CG
GLU
B
541
−92.860
−20.454
−19.911
1.00
32.56
C


ATOM
16555
CD
GLU
B
541
−93.773
−21.133
−20.935
1.00
33.64
C


ATOM
16556
OE1
GLU
B
541
−93.261
−21.922
−21.771
1.00
35.06
O


ATOM
16557
OE2
GLU
B
541
−95.002
−20.868
−20.901
1.00
32.44
O


ATOM
16558
C
GLU
B
541
−92.326
−23.510
−19.105
1.00
31.31
C


ATOM
16559
O
GLU
B
541
−92.860
−24.286
−18.310
1.00
31.30
O


ATOM
16562
MG
MG
C
1
−42.844
11.427
13.309
1.00
46.29
MG


ATOM
16561
MG
MG
C
2
−46.615
−18.454
−33.231
1.00
48.45
MG


ATOM
16563
O
HOH
E
1
−50.507
−5.408
−4.491
1.00
17.42
O


ATOM
16566
O
HOH
E
2
−64.748
−36.007
−11.725
1.00
2.00
O


ATOM
16569
O
HOH
E
3
−40.643
−2.220
−34.996
1.00
2.00
O


ATOM
16572
O
HOH
E
4
−36.090
−9.757
−37.074
1.00
17.20
O


ATOM
16575
O
HOH
E
5
−46.117
−37.662
−22.916
1.00
17.03
O


ATOM
16578
O
HOH
E
6
−49.541
35.476
6.921
1.00
14.03
O


ATOM
16581
O
HOH
E
7
−32.288
27.572
16.443
1.00
13.16
O


ATOM
16584
O
HOH
E
8
−50.706
10.207
−15.061
1.00
18.73
O


ATOM
16587
O
HOH
E
9
−77.188
36.767
−9.218
1.00
8.05
O


ATOM
16590
O
HOH
E
10
−90.260
−31.248
−21.071
1.00
2.00
O


ATOM
16593
O
HOH
E
11
−70.920
−33.414
−3.884
1.00
9.35
O


ATOM
16596
O
HOH
E
12
−37.761
−21.294
−9.249
1.00
25.78
O


ATOM
16599
O
HOH
E
13
−76.050
23.855
−18.293
1.00
2.00
O


ATOM
16602
O
HOH
E
14
−76.876
−19.575
−22.856
1.00
19.25
O


ATOM
16605
O
HOH
E
15
−40.936
11.629
−24.832
1.00
22.29
O


ATOM
16608
O
HOH
E
16
−85.551
34.832
3.260
1.00
25.14
O


ATOM
16611
O
HOH
E
17
−56.825
31.771
−7.464
1.00
17.27
O


ATOM
16614
O
HOH
E
18
−76.222
39.261
−.613
1.00
25.47
O









Various modifications and variations of the described method and system of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention, which are obvious to those skilled in the relevant fields, are intended to be within the scope of the following claims.

## Claims

1. An isolated host cell comprising a heterologous polynucleotide sequence encoding an isoprene synthase variant in operable combination with a promoter, wherein said isoprene synthase variant comprises one or more amino acid substitution(s) at one or more amino acid residues corresponding to a poplar isoprene synthase having the sequence of SEQ ID NO: 120, wherein said substitution(s) are selected from the group consisting of V10M, F12S, T15A, E18G, V58I, V58F, L70Q, L70R, L70V, L70T, T71P, V79L, E89D, G94A, S119F, F120L, G127R, E175V, T212I, S257A, R262G, A266G, F280L, N297K, F305L, L319M, E323K, A328T, D342E, A359T, K366N, E368D, L374M, S396T, V418S, K438N, H440R, T442A, I449V, A469S, K500R, K505Q, G507S, S509N, F511Y, and N532K; and
wherein the variant is capable of more effectively converting dimethylallyl diphosphate (DMAPP) to isoprene, as compared to an isoprene synthase variant without a substitution.

2. The host cell of claim 1 wherein at least one amino acid substitution is a L70R substitution.

3. The host cell of claim 1 wherein at least one amino acid substitution is a G507S substitution.

4. The host cell of claim 1 wherein the variant comprises one of more amino acid substitutions selected from the group consisting of G127R/F511Y, L70Q/G94A/R262G/F305L, F12S/T15A/E18G/N297K, S396T/T442I, V10M/E323K, F120L/A266G, K438N/K500R, V79L/S509N, E175V/S257A/E368D/A469S, T71P/L374M, F280L/H440R, E89D/H440R, V58F/A328T/N532K, S119F/D342E/I449V, and K366N/G507S.

5. The host cell of claim 1 wherein the polynucleotide sequence is contained within a plasmid.

6. The host cell of claim 5 wherein the polynucleotide sequence is integrated into a chromosome of the host cell.

7. The host cell of claim 1 wherein the host is selected from the group consisting of gram-positive bacterial cells, gram-negative bacterial cells, filamentous fungal cells, and yeast cells.

8. The host cell of claim 1 wherein the host is selected from the group consisting of Escherichia sp. (E. coli), Panteoa sp. (P. citrea), Bacillus sp. (B. subtilis), Saccharomyces sp. (S. cerevisia), Yarrowia sp. (Y. lipolytica), and Trichoderma (T. reesei).

9. The host cell of claim 1 wherein the host cell is cultured in a medium comprising a carbon source selected from the group consisting of glucose, glycerol, glycerine, dihydroxyacetone, yeast extract, biomass, molasses, sucrose, and oil.

10. The host cell of claim 1 wherein the host cell further comprises a heterologous or native nucleic acid encoding an isopentenyl-diphosphate delta-isomerase (IDI) polypeptide or a heterologous or native nucleic acid encoding a 1-Deoxyxylulose-5-phosphate synthase (DXS) polypeptide, optionally in combination with the native 1-deoxy-D-xylulose-5-phosphate (DXP) pathway.

11. The host cell of claim 1 wherein the host cell further comprises one or more nucleic acids encoding an IDI polypeptide and a DXS polypeptide.

12. The host cell of claim 1 wherein the host cell comprises one vector encoding the isoprene synthase variant, the IDI polypeptide, and the DXS polypeptide.

13. The host cell of claim 12 wherein the host cell further comprises a nucleic acid encoding a mevalonate (MVA) pathway polypeptide selected from the group consisting of an MVA pathway polypeptide from Saccharomyces cerevisia and Enterococcus faecalis.

14. The host cell of claim 1 wherein the host cell further comprises one or more nucleic acids encoding an MVA pathway polypeptide and a DXS polypeptide and wherein one vector encodes the isoprene synthase variant, the MVA pathway polypeptide, and the DXS polypeptide.

15. The host cell of claim 14 wherein the host cell further comprises one or more nucleic acids encoding a DXS polypeptide, an IDI polypeptide, or one or more of the rest of the DXP pathway polypeptides, and a MVA pathway polypeptide.

16. The host cell of claim 1 wherein the host cell further comprises one or more nucleic acid(s) encoding an MVA pathway polypeptide.

17. The host cell of claim 16 wherein the host cell further comprises one or more nucleic acids encoding an IDI polypeptide.

18. A method of producing isoprene, comprising:
(a) culturing the host cells of claim 1 under suitable culture conditions for production of isoprene; and
(b) producing the isoprene.

19. The method of claim 18 further comprising (c) recovering the isoprene.

20. The method of claim 19 further comprising (d) polymerizing isoprene.

21. The host cell of claim 1 wherein the host cell further comprises a heterologous or native nucleic acid encoding an isopentenyl-diposhpate delta-isomerase (IDI) polypeptide and a heterologous or native nucleic acid encoding a 1-Deoxyxylulose-5-phosphate synthase (DXS) polypeptide, optionally in combination with the native 1-deoxy-D-xylulose-5-phospahte (DXP) pathway.